Patent Grant
7109186
Human immunodeficiency virus (HIV) has been identified as the etiological agent responsible for acquired immune deficiency syndrome (AIDS), a fatal disease characterized by destruction of the immune system and the inability to fight off life threatening opportunistic infections. Recent statistics (UNAIDS: Report on the Global HIV/AIDS Epidemic, December 1998), indicate that as many as 33 million people worldwide are infected with the virus. In addition to the large number of individuals already infected, the virus continues to spread. Estimates from 1998 point to close to 6 million new infections in that year alone. In the same year there were approximately 2.5 million deaths associated with HIV and AIDS.
There are currently a number of antiviral drugs available to combat the infection. These drugs can be divided into three classes based on the viral protein they target and their mode of action. In particular, saquinavir, indinavir, ritonavir, nelfinavir and amprenavir are competitive inhibitors of the aspartyl protease expressed by HIV. Zidovudine, didanosine, stavudine, lamivudine, zalcitabine and abacavir are nucleoside reverse transcriptase inhibitors that behave as substrate mimics to halt viral cDNA synthesis. The non-nucleoside reverse transcriptase inhibitors, nevaripine, delavaridine and efavirenz inhibit the synthesis of viral cDNA via a non-competitive (or uncompetitive) mechanism. Used alone these drugs are effective in reducing viral replication. The effect is only temporary as the virus readily develops resistance to all known agents. However, combination therapy has proven very effective at both reducing virus and suppressing the emergence of resistance in a number of patients. In the US, where combination therapy is widely available, the number of HIV-related deaths has declined (Palella, F. J.; Delany, K. M.; Moorman, A. C.; Loveless, M. O.; Further, J.; Satten, G. A.; Aschman, D. J.; Holmberg, S. D. N. Engl. J. Med. 1998, 338, 853).
Unfortunately, not all patients are responsive and a large number fail this therapy. In fact, approximately 30–50% of patients ultimately fail combination therapy. Treatment failure in most cases is caused by the emergence of viral resistance. Viral resistance in turn is caused by the rapid turnover of HIV-1 during the course of infection combined with a high viral mutation rate. Under these circumstances incomplete viral suppression caused by insufficient drug potency, poor compliance to the complicated drug regiment as well as intrinsic pharmacological barriers to exposure provides fertile ground for resistance to emerge. More disturbing are recent findings which suggest that low-level replication continues even when viral plasma levels have dropped below detectable levels (<50 copies/ml) (Carpenter, C. C. J.; Cooper, D. A.; Fischl, M. A.; Gatell, J. M.; Gazzard, B. G.; Hammer, S. M.; Hirsch, M. S.; Jacobsen, D. M.; Katzenstein, D. A.; Montaner, J. S. G.; Richman, D. D.; Saag, M. S.; Schecter, M.; Schoolery, R. T.; Thompson, M. A.; Vella, S.; Yeni, P. G.; Volberding, P. A. JAMA 2000, 283, 381). Clearly there is a need for new antiviral agents, preferably targeting other viral enzymes to reduce the rate of resistance and suppress viral replication even further.
HIV expresses three enzymes, reverse transcriptase, an aspartyl protease and integrase, all of which are potential antiviral targets for the development of drugs for the treatment of AIDS. However, integrase stands out as being the only viral enzyme not targeted by current therapy. The integrase enzyme is responsible for insertion of the viral cDNA into the host cell genome, which is a critical step in the viral life cycle. There are a number of discrete steps involved in this process including processing of the viral cDNA by removal of two bases from each 3′-terminus and joining of the recessed ends to the host DNA. Studies have shown that in the absence of a functional integrase enzyme HIV is not infectious. Therefore, an inhibitor of integrase would be useful as a therapy for AIDS and HIV infection.
A number of inhibitors of the enzyme have been reported. These include, nucleotide-based inhibitors, known DNA binders, catechols and hydrazide containing derivatives (Neamati, N. Expert Opin. Ther. Patents 2002, 12, 709–724). Diketoamide HIV integrase inhibitors have been disclosed (WO 0316266, WO 0335076, WO 0335077, WO 02070486). However, no clinically approved compound has resulted from these leads. Thus, clinically effective inhibitors of HIV integrase would fulfill a therapeutic need.
The present invention relates to compounds of Formula I, or pharmaceutically acceptable salts and solvates thereof
wherein R1, R2, R3, and B1 are described as below. The invention includes compositions and methods of treatment using these compounds.
The present invention describes compounds of Formula I, or pharmaceutically acceptable salts or solvates thereof
wherein:
R1 is
R10 is
In the present invention, unless otherwise specified the following definitions apply.
The numbers in the subscript after the symbol “C” define the number of carbon atoms a particular group can contain. For example, “C1–C6” means a substituent containing from one to six carbon atoms.
As used herein, the term “alkyl” means a saturated, straight chain or branched monovalent hydrocarbon radical having the stated number of carbon atoms. Examples of such alkyl radicals include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, t-butyl and, where indicated, higher homologues and isomers such as n-pentyl, n-hexyl, 2-methylpentyl and the like. Haloalkyl refers to an alkyl radical that is substituted with one or more halo radicals, such as trifluoromethyl.
As used herein, the term “cycloalkyl” means a non-aromatic 3–6 membered ring. Examples include, cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.
Halo means chloro, bromo, iodo or fluoro.
“Aryl” means an aromatic hydrocarbon having from six to ten carbon atoms; examples include phenyl and napthyl, indenyl, azulenyl, fluorenyl and anthracenyl.
The term “heterocyle” refers to a monocyclic saturated heterocyclic nuclei having 3–6 atoms containing 1–3 heteroatoms selected from nitrogen, oxygen or sulfur. Heterocycles include, for example, piperidinyl, piperazinyl, pyrrolidinyl and morpholinyl.
“Heteroaryl” means a five- or six-membered aromatic ring containing at least one and up to four non-carbon atoms selected from oxygen, sulfur and nitrogen. Examples of heteroaryl include 2-furyl, 3-furyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, pyrazinyl, 2-thienyl, 3-thienyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, 1,2,3-oxadiazolyl, 1,2,3-triazolyl, 1,3,4-thiadiazolyl, pyridazinyl, pyrimidinyl, 1,3,5-triazinyl and 1,3,5-trithianyl.
By virtue of its acidic moiety, where applicable, a compound of Formula I forms salts by the addition of a pharmaceutically acceptable base. Such base addition salts include those derived from inorganic bases which include, for example, alkali metal salts (e.g. sodium and potassium), alkaline earth metal salts (e.g. calcium and magnesium), aluminum salts and ammonium salts. In addition, suitable base addition salts include salts of physiologically acceptable organic bases such as trimethylamine, triethylamine, morpholine, pyridine, piperidine, picoline, dicyclohexylamine, N,N′-dibenzylethylenediamine, 2-hydroxyethylamine, bis-(2-hydroxyethyl)amine, tri-(2-hydroxyethyl)amine, procaine, dibenzylpiperidine, N-benzyl-phenethylamine, dehydroabietylamine, N,N′-bishydroabietylamine, glucamine, N-methylglucamine, collidine, quinine, quinoline, ethylenediamine, ornithine, choline, N,N′-benzylphenethylamine, chloroprocaine, diethanolamine, diethylamine, piperazine, tris(hydroxymethyl)aminomethane and tetramethylammonium hydroxide and basic amino aids such as lysine, arginine and N-methylglutamine. These salts may be prepared by methods known to those skilled in the art.
Salts of an amine group may also comprise quaternary ammonium salts in which the amino nitrogen carries a suitable organic group such as an alkyl, alkenyl, alkynyl or aryl moiety.
Compounds of Formula I which are substituted with a basic group may exist as salts formed through acid addition. The acid addition salts are formed from a compound of Formula I and a pharmaceutically acceptable inorganic acid, including but not limited to hydrochloric, hydrobromic, hydroiodic, sulfuric, phosphoric, or an organic acid such as p-toluenesulfonic, methanesulfonic, acetic, benzoic, citric, malonic, fumaric, maleic, oxalic, succinic, sulfamic, or tartaric. Thus, examples of such pharmaceutically acceptable salts include chloride, bromide, iodide, sulfate, phosphate, methanesulfonate, citrate, acetate, malonate, fumarate, sulfamate, and tartrate.
Certain compounds of Formula I, and their salts, may also exist in the form of solvates with water, for example hydrates, or with organic solvents such as methanol, ethanol or acetonitrile to form, respectively, a methanolate, ethanolate or acetonitrilate. The present invention includes each solvate and mixtures thereof.
In addition, a compound of Formula I, or its salt or solvate, may exhibit polymorphism. The present invention also encompasses any such polymorphic form.
Certain compounds of Formula I may contain one or more chiral centers and exist in different optically active forms. When compounds of Formula I contain one chiral center, the compounds exist in two enantiomeric forms. The present invention includes both enantiomers and mixtures of enantiomers such as racemic mixtures. The enantiomers may be resolved by methods known to those skilled in the art, for example, by formation of diastereoisomeric salts which may be separated by crystallization, gas-liquid or liquid chromatography, selective reaction of one enantiomer with an enantiomer-specific reagent. It will be appreciated that where the desired enantiomer is converted into another chemical entity by a separation technique, then an additional step is required to form the desired enantiomeric form. Alternatively, specific enantiomers may be synthesized by asymmetric synthesis using optically active reagents, substrates, catalysts or solvents, or by converting one enantiomer into the other by asymmetric transformation.
Certain compounds of Formula I may also exist in different stable conformational forms which may be separable. Torsional asymmetry due to restricted rotation about an asymmetric single bond, for example because of steric hindrance or ring strain, may permit separation of different conformers. The present invention includes each conformational isomer of compounds of Formula I and mixtures thereof.
Certain compounds of Formula I may exist in zwitterionic form and the present invention includes each zwitterionic form of compounds of Formula I and mixtures thereof.
The compounds of this invention can also exist as tautomers; therefore the present invention also includes all tautomeric forms.
The compounds of Formula I are useful in the inhibition of HIV integrase, the prevention or treatment of infection by the human immunodeficiency virus and the treatment of consequent pathological conditions such as AIDS or ARC. The treatment involves administering to a patient, in need of such treatment, a compound of Formula I, or a pharmaceutically acceptable salt, solvate or prodrug thereof, or a pharmaceutical composition comprising a pharmaceutical carrier and a therapeutically effective amount of a compound of the present invention, or a pharmaceutically acceptable salt, solvate or prodrug therefor.
Treatment extends to prophylaxis as well as established infections or symptoms. This includes initiating treatment pre- and post-exposure to the virus. In addition, the present invention can be administered in conjunction with other anti-HIV agents (HIV protease inhibitors, nucleoside reverse transcriptase inhibitors, non-nucleoside reverse transcriptase inhibitors, and HIV-entry inhibitors), immunomodulators, antiinfectives and/or vaccines.
The compounds of the present invention are also useful in the preparation and execution of screening assays for antiviral compounds. Further, the compounds of the present invention are useful in establishing or determining the binding site of other antiviral compounds to HIV integrase, for example, by competitive inhibition.
The compounds of the present invention may be administered orally, parenterally (including subcutaneous injections, intravenous, intramuscular, intrasternal injection or infusion techniques), by inhalation spray or rectally, in dosage unit formulations containing conventional non-toxic pharmaceutically acceptable carriers, adjuvants and vehicles.
This invention also provides a pharmaceutical composition for use in the above described therapeutic method. A pharmaceutical composition of the present invention comprises an effective amount of a compound of Formula I in association with a pharmaceutically acceptable carrier, excipient or diluent.
The active ingredient in such formulations comprises from 0.1 percent to 99.9 percent by weight of the formulation. By “pharmaceutically acceptable” it is meant that the carrier, diluent or excipient must be compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.
The present pharmaceutical compositions are prepared by known procedures using well known and readily available ingredients. The compositions of this 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. In making the compositions of the present invention, the active ingredient will usually be admixed with a carrier, or diluted by a carrier, or enclosed within a carrier which may be in the form of a capsule, sachet, paper or other container. When the carrier serves as a diluent, it may be a solid, semi-solid or liquid material which acts as a vehicle, excipient or medium for the active ingredient. Thus, the compositions can be in the form of tablets, pills, powders, beadlets, lozenges, sachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols, (as a solid or in a liquid medium), soft and hard gelatin capsules, suppositories, sterile injectable solutions, sterile packaged powders and the like.
The compounds can be administered by a variety of routes including oral, rectal, transdermal, subcutaneous, intravenous, intramuscular and intranasal.
When administered orally, these compositions are prepared according to techniques well-known in the art of pharmaceutical formulation. For oral administration, the compound is typically formulated with excipients such as binders, fillers, lubricants, extenders, diluents, disintegration agents and the like as are known in the art.
For parenteral administration, the compound is formulated in pharmaceutically acceptable non-toxic, parenterally-acceptable diluents or solvents, such as mannitol, 1,3-butanediol, water, 5 percent dextrose, Ringer's solution or isotonic sodium chloride solution, or suitable dispersing or wetting and suspending agents, such as sterile, bland, fixed oils, including synthetic mono- or diglycerides, and fatty acids, including oleic acid.
A compound of the present invention, or a salt or solvate thereof, can be formulated in unit dosage formulations comprising a dose between about 0.1 mg and about 1000 mg, or more, according to the particular treatment involved. An example of a unit dosage formulation comprises 5 mg of a compound of the present invention in a 10 mL sterile glass ampoule. Another example of a unit dosage formulation comprises about 10 mg of a compound of the present invention as a pharmaceutically acceptable salt in 20 mL of isotonic saline contained in a sterile ampoule.
The compounds of the present invention can also be administered to humans in a dosage range of 1 to 100 mg/kg body weight in divided doses. One preferred dosage range is 1 to 20 mg/kg body weight orally in divided doses. It will be understood, however, that the specific dose level and frequency of dosage for any particular patient may be varied and will depend upon a variety of factors including the activity of the specific compound employed, the metabolic stability and length of action of that compound, the route of administration, the age, body weight, general health, sex, diet, mode and time of administration, rate of excretion, drug combination, the severity of the particular condition, and the host undergoing therapy.
General methods useful for the synthesis of compounds embodied in this invention are shown below. The preparations shown below are disclosed for the purpose of illustration and are not meant to be interpreted as limiting the processes to make the compounds by any other methods. It will be appreciated by those skilled in the art that a number of methods are available for the preparation of the compounds of the present invention as provided by Formula I.
Formula I compounds can be prepared by processes which include processes known in the chemical art for the production of structurally analogous compounds or by a novel process described herein. A process for the preparation of a compound of Formula I (or a pharmaceutically acceptable salt thereof) and novel intermediates for the manufacture of a compound of Formula I, as defined above, provide further features of the invention and are illustrated by the following procedures in which the meanings of the generic radicals are as defined above, unless otherwise specified. It will be recognized that it may be preferred or necessary to prepare a compound of Formula I in which a functional group is protected using a conventional protecting group, and then to remove the protecting group to provide the compound of Formula I.
Thus, there is provided a process for preparing a compound of Formula I (or a pharmaceutically acceptable salt thereof) as provided in any of the above descriptions which is selected from any of those described in the examples, including the following.
The compounds of the present invention can be synthesized according to the following schemes. Schemes I-III represent general methods for the synthesis of the compounds. In Scheme I, an appropriately substituted amine, I-1, can be acylated under standard amide bond forming conditions to yield I-3. Methods for this type of transformation are described, in Jerry March, Advanced Organic Chemistry, 3rd edition, John Wiley & Sons, 1985. The acetylamide I-3 can be condensed with dimethyl oxalate in the presence of a base such as NaOMe or LiHMDS. In the final step of the sequence, I-5 can be treated with and aldehyde, I-6, and an amine, I-7, to deliver the desired product I-8.
Scheme II illustrates an alternative synthesis. In this route, amine I-1 can be coupled to II-1 using standard amide bond forming methods. The product of this reaction is II-2 which can be converted into I-5 by methanolysis of the dioxalane ring system and the resulting product carried on to the final product in a manner similar to that in Scheme I. In scheme III, compound II-2 can be synthesized as before, but instead of forming intermediate I-5 it can be converted directly to I-8 according to the equation.
In Scheme IV, a substituted benzoic acid derivative can be coupled with amine IV-2 using standard amide bond forming methods to yield IV-3. This intermediate can be reduced to the corresponding benzylic amine, IV-4, which can be coupled to I-2. Intermediate IV-5 can then be condensed with dimethyl oxalate under basic conditions resulting in ketoacid IV-6. The final product can be delivered by treating IV-6 with paraformaldehyde and amine I-7 in acetic acid at elevated temperature.
In Scheme V, (3,4-dichloro-benzyl)-methylamine, V-1, can be coupled to III-B to yield Compound 37-A. This compound can then be treated with paraformaldehyde and amine I-7 resulting in V-2.
In Scheme VI, ketone or aldehyde, VI-1, can be condensed with an alkyl-hydroxylamine, VI-2, to yield the corresponding oxime. This can be then reduced with NaBH3CN to amine VI-4. It will be appreciated by those skilled in the art that this reduction can be carried out with a number of different reducing agents. Intermediate VI-4 can be coupled with III-B to yield VI-5 which can be taken on to VI-7 by two alternative routes. In one, VI-5 can be first treated with methanol to yield the corresponding methyl ester, VI-6, which can be converted to VI-7 as described previously. In the alternative procedure VI-5 can be treated with paraformaldehyde and I-7 as described in Scheme III.
In Scheme VII, amine VII-2 can be attached to an aldehyde-functionalized polystyrene resin (4-formyl-3-methoxy-phenoxymethyl functionalized polystyrene), VII-1, via reductive amination using methodology well known in the art. This intermediate can be coupled to acid, III-A using standard amide bond forming reaction conditions. Intermediate VII-4 can be treated with I-6 and I-7 to yield VII-5. The final product can be cleaved from the resin under acidic conditions to yield product VII-6.
In Scheme VIII, intermediate carboxylic acid, VIII-1 can be converted to the corresponding acid chloride, VIII-2, under standard conditions. This compound can then be treated with amine VIII-3 under basic conditions to yield amide VIII-4. Amide VIII-4 can be treated with paraformaldehyde and methylamine to provide the final product, VIII-5.
In Scheme IX, compound IX-1 can be synthesized according to the procedure of Heynes R. et al. Bull. Soc. Chim. Fr. (1977) 906–910 and reacted with CH3I or Ac2O to yield intermediates IX-2 and IX-5 respectively. Saponification or hydrogenolysis of the benzyl ester can provide carboxylic acids IX-3 and IX-6 which can be coupled with I-1 under amide bond forming reaction conditions. In the final step of the synthesis the methyl enol of XI-4 and the acetyl enol of IX-7 can be removed to deliver the final product IX-8.
In scheme X, Compound 82-A can be treated with trifluoroacetic acid to effect hydrolysis of the dimethyl-acetal. This then can be reacted with an heterocycle X-1 and a reducing agent such as sodium cyanoborohydride (NaBH3CN) to yield X-2. It will be understood by those skilled in the art that alternative reducing agents exist which can be used to carry out the same transformation.
In scheme XI Compound 24 can be reacted with amines, XI-1 and XI-3, using standard amide bond forming reagents to form compounds XI-2 and XI-4 respectively.
Another method for the synthesis of compounds of the current invention is illustrated in scheme XII. In this scheme Compound 84-A can be reacted with the corresponding acid chloride XII-1, sulfamoyl chloride XII-3 or sulfonyl chloride XII-5 under basic conditions to deliver compounds XII-2, XII-4 and XII-6 respectively.
In still another method to synthesize compounds of formula V-2, amine I-7 and paraformaldehyde can be reacted in methanol at elevated temperature to form intermediate XIII-1 as shown in scheme XIII. This intermediate is not isolated but added to a methanolic solution of Compound 37-A to yield compounds of formula V-2.
The synthesis of compounds of formula XVI-1 is illustrated in Scheme XIV. In this scheme Compound 93-A can be converted to the corresponding methyl ester, 93-B, as described in the schemes above. Amine I-7 and paraformaldehyde can be condensed to form intermediate XIII-1 which is reacted with 93-B as before to yield XIV-1.
Compounds of this invention can also be synthesized according to the method illustrated in Scheme XV. In this method 2,4-difluorobenzaldehyde can be treated with thiomethoxide to generate 2-methylthio-4-fluorobenzaldehyde which can be converted to compound XV-1 via reductive amination with amine R3NH2 (IV-2). This intermediate in turn can be acylated with III-B to yield XV-2. Oxidation of the sulfide to the corresponding sulfoxide and sulfone can be carried out under conditions familiar to those skilled in the art. Conversion of XV-3 and XV-4 to the corresponding compounds of formulas XV-5 and XV-6 can be achieved by reaction with amine I-7 and paraformaldehyde as described in previous methods.
In another method Compound 120-B can be treated with I-7 and paraformaldehyde to yield compounds of formula XVI-1, as illustrated in Scheme XVI.
The specific examples that follow illustrate the syntheses of the compounds of the instant invention, and are not to be construed as limiting the invention in sphere or scope. The methods may be adapted to variations in order to produce compounds embraced by this invention but not specifically disclosed. Further, variations of the methods to produce the same compounds in somewhat different manner will also be evident to one skilled in the art.
In the following experimental procedures, all temperatures are understood to be in Centigrade (C) when not specified. The nuclear magnetic resonance (NMR) spectral characteristics refer to chemical shifts (δ) expressed in parts per million (ppm) versus tetramethylsilane (TMS) as reference standard. The relative area reported for the various shifts in the proton NMR spectral data corresponds to the number of hydrogen atoms of a particular functional type in the molecule. In certain cases where the product is isolated as an inseparable mixture of isomers the integration of protons is given in decimal fractions corresponding to the proportion of that particular isomer's protons in the mixture. The nature of the shifts as to multiplicity is reported as broad singlet (bs or br s), broad doublet (bd or br d), broad triplet (bt or br t), broad quartet (bq or br q), singlet (s), multiplet (m), doublet (d), quartet (q), triplet (t), doublet of doublet (dd), doublet of triplet (dt), and doublet of quartet (dq). The solvents employed for taking NMR spectra are acetone-d6 (deuterated acetone), DMSO-d6 (perdeuterodimethylsulfoxide), D2O (deuterated water), CDCl3 (deuterochloroform) and other conventional deuterated solvents.
The abbreviations used herein are conventional abbreviations widely employed in the art. Some of which are: calcd (calculated); DMSO (dimethylsulfoxide); EDC (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride); EtOAc (ethyl acetate); HOBt (1-hydroxybenzotriazole); HPLC (high-pressure liquid chromatography); LC/MS (liquid chromatography, mass spectroscopy); LDA (lithium diisopropyl amide); LiHMDS (lithium bis(trimethylsilyl)amide); MCPBA (3-chloroperoxybenzoic acid) SiO2 (silica gel); THF (tetrahydrofuran), TFA (trifluoroacetic acid), Me (methyl), Et (ethyl), Ph (phenyl), tBuOK (potassium tert-butoxide), NaOMe (sodium methoxide), NaOEt (sodium ethoxide), Boc (tert-butoxycarbonyl), and DEAD (diethylazo dicarboxylate).
A solution of (S)-(+)-2,2-dimethyl-5-oxo-1,3-dioxolane-4-acetic acid (2.08 g, 11.9 mmol) in dry dichloromethane (20 ml) was treated with triethylamine (1.83 ml, 13.1 mmol) followed by a solution of t-butylchlorodiphenylsilane (3.44 g, 12.5 mmol) in dichloromethane (5 ml) added dropwise over 5 minutes. After 3 hours at 22° C., the reaction mixture was diluted with toluene (250 ml) washed with water, saturated sodium bicarbonate, brine and dried over magnesium sulfate. Evaporation of the solvent under reduced pressure and chromatography of the residue on silica gel (4×12 cm) using a mixture of toluene and ethyl acetate (0–2%) as eluent gave 4.90 g (99% yield) of the title material as a clear oil. 1H NMR (400 MHz, CDCl3) δ: 1.13 (s, 9), 1.58 (s, 3), 3.05 (m, 2), 4.79 (dd, 1, J=4, 7), 7.4–7.8 (m, 10).
A solution of (S)-(+)-2,2-dimethyl-5-oxo-1,3-dioxolane-4-acetic acid, tert-butyldiphenylsilyl ester (21.65 g, 52.4 mmol) in carbon tetrachloride (160 ml) was treated with N-bromosuccinimide (9.35 g, 52.4 mmol) and 2,2′-azobisisobutyronitrile (200 mg) and the resulting mixture was heated under reflux (bath temperature 85° C.) while irradiating with a 500 watt lamp. After 10 minutes, the reaction mixture was cooled and the succinimide was filtered. The solvent was evaporated under vacuum to give the title bromide as a light yellow oil (˜26 g) which was used immediately for the next step. 1H NMR (400 MHz, CDCl3) δ: 1.12 (s, 9), 1.41 (s, 3), 1.80 (s, 3), 3.80 (m, 2), 7.3–7.7 (m, 10).
A solution of 4-bromo-2,2-dimethyl-5-oxo-1,3-dioxolane-4-acetic acid, tert-butyldiphenylsilyl ester (˜26 g, 52.4 mmol) in dry tetrahydrofuran (160 ml) was cooled to 0° C. and treated dropwise over 5 minutes with 1,8-diazabicyclo[5,4,0]undec-7-ene (12.7 g, 78.8 mmol) and the resulting mixture was stirred at 5° C. for 1.5 hour. The solid formed was filtered and washed with a small amount of tetrahydrofuran. The filtrate was used as such for the next step.
Alternatively, the reaction mixture can be diluted with toluene, washed with water, saturated sodium bicarbonate, brine and dried (magnesium sulfate). Evaporation of the solvent gave an oil which was chromatographed on silica gel using a mixture of toluene and ethyl acetate (0–2%) as eluent. The title ester was obtained as an oil in 30–50% yield. 1HNMR (400 MHz, CDCl3) δ: 1.16 (s, 9), 1.76 (s, 6), 5.97 (s, 1), 7.4–7.8 (m, 10).
A solution of pure (Z)-2,2 dimethyl-5-(t-butyldiphenylsilyloxy-carbonylmethylene)-1,3-dioxolan-4-one (2.80 g, 6.82 mmol) in tetrahydrofuran (40 ml) was treated at 22° C. with acetic acid (2 ml) followed by 6.8 ml of a 1 M solution of tetrabutylammonium fluoride in tetrahydrofuran. After 15 minutes at 22° C., the reaction mixture was diluted with ethyl acetate, washed with water, brine and dried (magnesium sulfate). The solvent was concentrated under reduced pressure and the residue was triturated with toluene to give 1.00 g (85% yield) of the title compound as a white crystalline material: mp 203–204° C. (dec.). IR (KBr) ν max (cm−1): 1805, 1707 and 1662. 1H NMR (400 MHz, CDCl3) δ: 1.78 (s, 6), 5.89 (s, 1). Anal. calcd for C7H8O5: C, 48.84; H, 4.68. Found: C, 48.84; H, 4.65.
A solution of the crude (Z)-2,2-dimethyl-5-(tert-butyldiphenylsilyloxycarbonyl methylene)-1,3-dioxolan-4-one (52.4 mmol) in tetrahydrofuran (200 ml) was treated with acetic acid (13 ml) followed with 50 ml of a 1 M solution of tetrabutylammonium fluoride in tetrahydrofuran. After 15 minutes at 22° C., the reaction mixture was filtered and the filtrate was concentrated in vacuo. Trituration of the residue with toluene gave 6.3 g (70% yield for three steps) of the title material as a white solid (>95% pure by 1HNMR).
A solution of (S)-(+)-2,2-dimethyl-5-oxo-1,3-dioxolane-4-acetic acid (13.20 g, 75.8 mmol) in N,N-dimethylformamide (25 ml) was treated at 22° C. with imidazole (10.56 g, 0.155 mmol) followed by tert-butyldimethylsilyl chloride (12.0 g, 79.6 mmol) and the resulting mixture was stirred at 22° C. for 18 hours. The reaction mixture was then diluted with toluene (500 ml), washed with water (×3), saturated sodium bicarbonate and brine. After drying (magnesium sulfate), the solvent was evaporated under reduced pressure to give an oil. Distillation under vacuum gave 20.9 g (96% yield) of the title material as a clear oil: Bp 80–90° C./0.1 torr (bulb to bulb distillation, air bath temperature). 1H NMR (400 MHz, CDCl3) δ: 0.33 (s, 3), 0.36 (s, 3), 1.00 (s, 9), 1.11 (s, 3), 1.37 (s, 3), 2.72 (m, 2), 4.35 (dd, 1, J=4, 6).
A solution of (S)-(+)-2,2-dimethyl-5-oxo-1,3-dioxolane-4-acetic acid, t-butyldimethylsilyl ester (20.9 g, 72.4 mmol) in carbon tetrachloride (200 ml) was treated with N-bromosuccinimide (14.18 g, 79.6 mmol) and 2,2′-azobisisobutyronitrile (0.30 g) and the resulting mixture was heated under reflux while irradiating with a 500 W lamp. After approximately 5 minutes, a mild exothermic reaction was observed and the mixture was heated for an additional 5 minutes. The reaction mixture was then cooled in an ice bath and the floating succinimide was filtered and washed with a small amount of carbon tetrachloride. The filtrate was used immediately as such for the next step. 1H NMR (400 MHz, CDCl3) δ: 0.27 (s, 3), 0.28 (s, 3), 0.94 (s, 9), 1.66 (s, 3), 1.84 (s, 3), 3.62 (m, 2).
The solution of crude 4-bromo-2,2-dimethyl-5-oxo-1,3-dioxolane-4-acetic acid, tert-butyldimethylsilyl ester (72.4 mmol) in carbon tetrachloride (approximately 220 ml) was cooled to 0–5° C. and treated dropwise over 10 minutes and under vigorous stirring with a solution of 1,8-diazabicyclo[5,4,0]undec-7-ene (12.1 g, 79.6 mmol) in dry tetrahydrofuran (125 ml). A heavy precipitate was formed which gradually became a granular solid. After 1 h, the solid obtained was filtered and washed with a small amount of tetrahydrofuran. The filtrate was concentrated under reduced pressure to give a light orange oil which was used as such for the next step.
The crude (Z)-2,2-dimethyl-5-(tert-butyldimethylsilyloxycarbonyl-methylene)-1,3-dioxolan-4-one (72.4 mmol) in tetrahydrofuran (50 ml) was treated at 22° C. with acetic acid (13 ml, 0.227 mmol) followed by 73 ml (73.0 mmol) of a 1M solution of tetrabutylammonium fluoride in tetrahydrofuran. After 1 h at 22° C., the reaction mixture was diluted with ethyl acetate (500 ml), washed with water, brine and dried (anhydrous magnesium sulfate). Evaporation of the solvent under reduced pressure and trituration of the residual solid with toluene (50 ml) gave 7.70 g (62% yield for 3 steps) of the title Z-isomer as a white crystalline solid. Concentration of the mother liquors yielded another 0.2 g of a 75:25 mixture of Z and E isomers. Z-Isomer; 1H NMR (400 MHz, CDCl3) δ: 1.78 (s, 3), 5.89 (s, 1). E-Isomer: 1H NMR (400 MHz, CDCl3) δ: 1.80 (s, 3), 6.03 (s, 1).
A mixture of (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetic acid (0.50 g, 2.9 mmol) in dry dichloromethane (10 ml) was treated at 22° C. with oxalyl chloride (0.5 ml, 5.8 mmol) followed by a trace (capillary) of N,N-dimethylformamide. After 1 h at 22° C., the clear solution was concentrated in vacuo to give 0.55 g (quantitative) of the title acid chloride as a white crystalline solid.
To a solution of 30 mL MeNH2 (40% wt in H2O) was added 139 mL of 1N NaOH. To this was added 130 mL of CH2Cl2 followed by 4-fluorobenzoyl chloride (22 grams, 139 mmol) while the mixture was rapidly stirred. After 1 hour the organic layer was separated, washed with H2O, dried over Na2SO4 and solvent removed under vacuum to yield 20 grams (94% yield) solid. 1H NMR (500 MHz, DMSO) δ: 2.78 (d, 3, J=5), 7.28 (t, 2, J=9), 7.91 (m, 2), 8.46 (br s, 1). 13C NMR (125 MHz, DMSO) δ: 26.13, 114.95, 115.13, 129.49, 129.56, 130.87, 130.90, 162.67, 164.64, 165.46.
4-Fluoro-N-methyl-benzamide (18.8 grams, 123 mmol) was dissolved in 180 mL of THF. To this was added BF3.Et2O (5.51 mL, 43 mmol) and the resulting mixture heated to reflux for 15 min. The solution was then cooled to −20° C. and BH3.S(CH3)2 (16.5 mL, 174 mmol) added over 10 min. After this the reaction mixture was heated and the solvent removed by distillation for 20 min. The distillation apparatus was replaced with a reflux condenser and the reaction heated to 110° C. for 2 h. After cooling to room temperature 75 mL of 6N HCl was slowly added. After gas evolution had ceased the mixture was heated at reflux for 1 h then allowed to regain room temperature. To this was added 200 mL of 6N NaOH. The mixture was extracted with Et2O. The organic layer was washed with saturated NaCl, dried over Na2SO4, filtered and the solvent removed to yield an oil. The oil was dissolved in Et2O and 30 mL of 4N HCl (dioxane) added resulting in a white ppt. which was filtered to yield 18.9 grams solid (88% yield). 1H NMR (500 MHz, DMSO) δ: 2.48 (s, 3), 4.08 (s, 2), 7.26 (m, 2), 7.63 (m, 2), 9.60 (br s, 2). 13C NMR (125 MHz, DMSO) δ: 31.57, 50.04, 115.24, 115.41, 128.29, 128.32, 132.29, 132.36, 161.24, 163.19.
4-Fluoro-benzyl)-methyl-amine; hydrochloride (8.75 grams, 50 mmol) was added to a rapidly stirring mixture of 100 mL CH2Cl2 and 150 mL of 1N NaOH. To this was added acetyl chloride (3.55 mL, 50 mmol) and the resulting mixture stirred overnight. The organic layer was then separated, washed with 1N HCl, dried over Na2SO4, filtered and the solvent removed to yield 2 grams oil (22% yield). HRMS (M+H) calcd for C10H14FNO: 182.0891; found: 182.0979. 1H NMR (500 MHz, DMSO) δ: 2.04 (s), 2.05 (s), 2.77 (s), 2.90 (s), 4.46 (s), 4.53 (s), 7.13–7.28 (m, 4). 13C NMR (125 MHz, DMSO) δ: 21.16, 21.47, 32.85, 35.24, 48.81, 52.40, 114.97, 115.14, 115.30, 115.47, 128.52, 128.59, 129.38, 128.44, 133.58, 133.60, 134.01, 134.03, 160.23, 160.33, 162.16, 162.26, 169.70, 169.84.
N-(4-Fluoro-benzyl)-N-methyl-acetamide (3.6 grams, 20 mmol) was dissolved in 20 mL of THF and cooled to −78° C. To this was added 40 mL of 1M LiHMDS (in THF) and the resulting solution stirred for 30 min. Next dimethyl oxalate (3.5 grams, 30 mmol) dissolved in 8 mL of THF is added and the reaction stirred for 2 h at −78° C., then warmed to 0° C. and stirred an additional 30 min. To this was added 1N HCl and the mixture then extracted with EtOAc. The organic layer was washed with saturated NaCl, dried over Na2SO4, filtered and the solvent removed. The crude product was purified by column chromatography (4×4 cm SiO2, 80:20 Hex/EtOAc) to yield 4.3 grams solid (80% yield). HRMS (M+H) calcd for C13H15NO4F: 268.0985; found: 268.0983. Anal calcd for C13H14NO4F: C, 58.42; H, 5.28; N, 5.24. found: C, 58.48; H, 5.21; N, 5.26. 1H NMR and 13C NMR show a mixture of rotamers at room temperature. 1H NMR (500 MHz, CDCl3) δ: 3.00 (s), 3.86 (s), 3.89 (s), 4.55 (s), 6.29 (s), 6.31 (s), 7.00–7.24 (overlapping m, 4). 13C NMR δ; 33.43, 34.79, 49.97, 52.63, 52.97, 93.27, 93.55, 115.63, 115.80, 115.95, 116.13, 128.36, 128.42, 129.71, 129.78, 131.32, 132.03, 159.70, 161.40, 163.25, 163.35, 170.93, 171.16.
To 1.6 mL of ethanol at 60° C. was added paraformaldehyde (46.6 mg, 1.6 mmol. eq. of formaldehyde) and 0.78 mL of 2 M (THF) CH3NH2 and the resulting mixture stirred for 5 min. Compound 1-D (41.5 mg, 1.6 mmol) was added and the reaction stirred for 1 hr. The reaction mixture was then diluted with H2O and extracted with EtOAc. The organic layer was separated, washed with satd NaCl, dried over Na2SO4, filtered and the solvent removed to yield crude product, which was purified by preparative TLC (SiO2, 20% EtOH/CH2Cl2). HRMS [M+H] calcd for C14H16FN2O3: 279.1145. Found: 279.1148. 1H NMR (CDCl3, 500 MHz) δ: 2.99 (s, 3), 3.03 (s, 3), 4.13 (s, 2), 4.60 (s, 2), 6.98–7.23 (overlapping m, 4).
3-[(4-Fluoro-benzyl)-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 1-D) was treated with paraformaldehyde and 2-(4-chlorophenyl)-ethylamine as described in the preparation of Compound 1. HRMS (M−H) calcd for C21H19ClFN2O3; 401.1068. found: 401.1080. 1H NMR (500 MHz, CDCl3)δ: 2.90 (t, 2, j=7), 2.95 (s, 3), 3.72 (t, 2, J=7), 3.95 (s, 2), 4.57 (s, 2), 7.03–7.26 (overlapping m, 8). 13C NMR (125 MHz, CDCl3) δ: 33.92, 34.45, 44.42, 49.37, 51.46, 108.49, 118.91, 128.87, 129.22, 129.98, 131.99, 132.67, 136.60, 154.21, 161.42, 163.37, 164.51, 166.31.
3-[(4-Fluoro-benzyl)-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 1-D) was treated with paraformaldehyde and 2-(2-chlorophenyl)-ethylamine as described in the preparation of Compound 1. HRMS (M+H) calcd for C21H21ClFN2O3: 403.1225. found: 403.1237. 1H NMR (500 MHz, CDCl3) δ: 2.96 (s, 3), 3.05 (t, 2, J=7), 3.75 (t, 2, J=7), 3.99 (s, 2), 4.58 (s, 2), 7.01–7.34 (overlapping m, 8). 13C NMR (125 MHz, CDCl3) δ: 32.37, 34.52, 42.90, 49.36, 51.47, 109.38, 115.66, 115.83, 127.18, 128.37, 129.26, 129.32, 129.67, 130.92, 132.13, 132.15, 134.01, 135.79, 152.71, 161.37, 163.33, 164.91, 166.11.
3-[(4-Fluoro-benzyl)-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 1-D) was treated with paraformaldehyde and 2-thiophen-2-yl-ethylamine as described in the preparation of Compound 1. HRMS (M−H) calcd for C19H18FN2O3S: 373.1022. found: 373.1029. 1H NMR (500 MHz, CDCl3) δ: 2.95 (s, 3), 3.14 (t, 2, J=7), 3.76 (t, 2, J=7), 3.97 (s, 2), 4.57 (s, 2), 6.81–7.21 (overlapping m, 7). 13C NMR (125 MHz, CDCl3) δ: 28.76, 34.49, 44.93, 49.46, 51.47, 109.48, 115.65, 115.82, 124.20, 125.59, 127.15, 129.28, 129.34, 132.12, 132.14, 140.34, 152.72, 161.37, 163.33, 164.94, 166.11.
3-[(4-Fluoro-benzyl)-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 1-D) was treated with paraformaldehyde and 2-(2-fluoro-phenyl)-ethylamine as described in the preparation of Compound 1. HRMS (M+H) calcd for C21H21N2F2O3: 387.1520. found: 387.1525. 1H NMR (500 MHz, CDCl3) δ: 2.96 (overlapping m, 5), 3.75 (t, 2, J=7), 4.02 (s, 2), 4.58 (s, 2), 6.98–7.22 (overlapping m, 8). 13C NMR (125 MHz, CDCl3) δ: 28.13, 28.14, 34.55, 43.38, 49.34, 51.52, 109.53, 115.31, 115.48, 115.68, 115.85, 124.38, 124.41, 124.87, 124.99, 128.68, 128.75, 129.29, 129.35, 130.96, 130.99, 131.98, 132.01, 152.46, 160.26, 161.40, 162.21, 163.36, 165.04, 166.09.
3-[(4-Fluoro-benzyl)-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 1-D) was treated with paraformaldehyde and 2-(2,4-dichloro-phenyl)-ethylamine as described in the preparation of Compound 1. 1H NMR (500 MHz, CDCl3) δ: 2.99 (s, 3), 3.03 (t, 2, J=7), 3.74 (t, J=7), 4.06 (s, 2), 4.60 (s, 2), 7.03–7.37 (overlapping m, 7). 13C NMR (125 MHz, CDCl3) δ: 31.76, 34.67, 42.95, 49.48, 51.61, 109.74, 115.76, 115.93, 127.53, 129.27, 129.32, 129.53, 131.63, 131.78, 131.80, 133.57, 134.14, 134.65, 152.19, 161.44, 163.40, 165.24, 165.98.
3-[(4-Fluoro-benzyl)-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 1-D) was treated with paraformaldehyde and 2-pyridin-4-yl-ethylamine as described in the preparation of Compound 1. HRMS (M+H) calcd for C20H21FN3O3: 370.1567. found: 370.1568. 1H NMR (500 MHz, CDCl3) δ: 3.00 (s, 3), 3.23 (t, 2, J=7), 3.90 (t, 2, J=7), 4.20 (s, 2), 4.60 (s, 2), 7.03 (m, 2), 7.21 (m, 2), 7.80 (d, 2, J=6), 8.73 (d, 2, J=6). 13C NMR (125 MHz, CDCl3) δ: 34.55, 34.63, 42.47, 49.04, 51.55, 109.53, 114.58, 115.74, 115.91, 127.10, 129.30, 129.50, 131.76, 141.92, 153.22, 158.36, 160.92, 161.23, 161.43, 163.40, 165.33, 166.13.
3-[(4-Fluoro-benzyl)-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 1-D) was treated with paraformaldehyde and 2-phenyl-ethylamine as described in the preparation of Compound 1. HRMS (M+H) calcd for C21H22FN2O3: 369.1615. found: 369.1625. 1H NMR (500 MHz, CDCl3) δ: 2.92 (overlapping m, 5), 3.74 (t, 2, J=7), 3.92 (s, 2), 4.56 (s, 2), 7.01–7.29 (overlapping m, 9). 13C NMR (125 MHz, CDCl3) δ: 34.45, 34.60, 44.70, 49.42, 51.46, 109.12, 115.66, 115.83, 126.75, 128.63, 128.72, 129.26, 129.33, 132.09, 132.11, 138.18, 153.11, 161.38, 163.34, 164.79, 166.18.
3-[(4-Fluoro-benzyl)-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 1-D) was treated with paraformaldehyde and 2-(4-fluoro-phenyl)-ethylamine as described in the preparation of Compound 1. (HRMS (M−H) calcd for C21H19F2N2O3: 385.1364. found: 385.1377. 1H NMR (500 MHz, CDCl3) δ: 2.90 (t, 2, J=7), 2.95 (s, 3), 3.71 (t, 2, J=7), 3.95 (s, 2), 4.57 (s, 2), 6.95–7.26 (overlapping m, 8). 13C NMR (125 MHz, CDCl3) δ: 33.74, 34.48, 44.70, 49.41, 51.47, 108.82, 115.49, 115.66, 115.71, 115.77, 115.89, 129.22, 129.27, 130.04, 130.10, 131.97, 133.72, 153.60, 160.81, 161.42, 162.76, 163.38, 164.71, 166.21.
3-[(4-Fluoro-benzyl)-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 1-D) was treated with paraformaldehyde and 3-phenyl-propylamine as described in the preparation of Compound 1. 1H NMR (500 MHz, CDCl3) δ: 1.93 (p, 2, J=7), 2.64 (t, 2, J=7), 2.99 (s, 3), 3.53 (t, 2, J=2), 4.10 (s, 2), 4.60 (s, 2), 7.02–7.28 (overlapping m, 9). 13C (125 MHz, CDCl3) δ: 29.77, 33.05, 34.56, 42.81, 48.79, 51.47, 109.25, 115.66, 115.83, 126.14, 128.30, 128.49, 129.34, 129.40, 132.16, 132.79, 140.88, 152.46, 161.39, 163.34, 165.02, 166.08.
3-[(4-Fluoro-benzyl)-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (compound 1-D) was treated with paraformaldehyde and isopropylamine as described in the preparation of Compound 1. 1H NMR (500 MHz, CDCl3) δ: 1.23 (d, 6, J=7), 3.02 (s, 3), 4.10 (s, 2), 4.45 (heptet, 1, J=7), 4.62 (s, 2), 7.03 (m, 2), 7.24 (m, 2). 13C NMR (125 MHz, CDCl3) δ: 20.61, 34.72, 44.09, 44.34, 110.21, 115.57, 115.75, 129.43, 129.50, 132.28, 132.30, 149.92, 161.35, 163.31, 164.76, 165.80.
A solution of (3,4-dichlorobenzyl)-methylamine (0.50 g, 2.63 mmol) (Shapiro et al. J. Amer. Chem. Soc., (1959) 81, 3725) in a mixture of tetrahydrofuran (20 ml) and 40% aqueous sodium acetate (10 ml) was cooled to 0–5° C. (ice bath) and treated with a solution of acetyl chloride (0.3 g in tetrahydrofuran) added dropwise over 5 min. After 1 h at 0–5° C., the reaction mixture was diluted with ethyl acetate, washed successively with 1 N hydrochloric acid, saturated sodium bicarbonate, brine and dried (magnesium sulfate). Evaporation of the solvent and distillation of the residue in vacuo gave 0.51 g (83% yield) of the title amide as a clear oil: bp 110–120° C./0.2 torr, (bulb to bulb distillation, air bath temperature). 1HNMR 400 MHz (CDCl3) δ (ppm), mixture of rotamers: 2.36 and 2.39 (3H, 2 s, COCH3), 3.16 (3H, s, NCH3), 4.70 and 4.75 (2H, 2 s, NCH2), 7.2–7.7 (3H, m, aromatics). Anal. calcd for C10H11Cl2NO: C, 51.74; H, 4.78; N, 6.03. Found: C, 51.70; H, 4.77; N, 6.04.
A solution of N-(3,4-dichlorobenzyl)-N-methyl-acetamide (0.83 g, 3.57 mmol) in tetrahydrofuran (15 ml) was cooled to −78° C. and treated dropwise with 7.1 ml (7.1 mmol) of 1M lithium bis(trimethylsilyl)amide in tetrahydrofuran. After 20 min, the mixture was treated dropwise with a solution of dimethyl oxalate (0.63 g, 5.35 mmol) in tetrahydrofuran (3 ml), stirred at −78° C. for 1 h and then at 5° C. for another 45 min. The reaction mixture was then quenched by the addition of 1 N hydrochloric acid and ethyl acetate. The organic phase was washed successively with water, saturated sodium bicarbonate and brine and dried over anhydrous sodium sulfate. Evaporation of the solvent under reduced pressure and chromatography of the residue on silica gel (elution toluene-ethyl acetate, 85:15) gave 0.70 g (61% yield) of the title ester as clear oil. 1HNMR 400 MHz (C6D6) δ (ppm); mixture of rotamers: 2.01 and 2.49 (3H, 2 s, NCH3), 3.43 and 3.5 (3H, 2 s, OCH3), 4.0 and 4.4 (2H, 2 s, NCH2), 6.26 and 6.33 (1H, 2 s, CH), 6.62–7.2 (3H, m, aromatics). Anal. calcd for C13H13Cl2NO4: C, 49.08; H, 4.12; N, 4.40. Found: C, 49.38; H, 4.23; N, 4.30.
A solution of 2 M methylamine in tetrahydrofuran (0.5 ml, 1.0 mmol) was added to a mixture of paraformaldehyde (0.3 g, 1.0 mmol, equivalent of formaldehyde) in anhydrous ethanol (1 ml) and the resulting mixture was heated at 60° C. for 5 min. Then a solution of 3-[(3,4-dichlorobenzyl)-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (0.318 g, 1.0 mmol) in ethanol (3 ml) was added all at once and the resulting mixture was maintained at 60° C. for another 20 min. The reaction mixture was then quenched by the addition of ethyl acetate and pH 2 phosphate buffer. The organic phase was washed with brine, dried (anhydrous magnesium sulfate) and concentrated under reduced pressure. Recrystallization of the solid residue from a mixture of ethyl acetate and hexane gave 0.180 g (54% yield) of the title material as a white solid: mp 148–150° C.
To 0.5 mL of AcOH at 60° C. was added N-(2-aminoethyl)morpholine (0.066 mL, 0.5 mmol) and paraformaldehyde (15 mg, 0.5 mmol). After stirring for 5 min, 3-[(3,4-dichloro-benzyl)-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (159 mg, 0.5 mmol) was added and the resulting mixture stirred at 60° C. for 2 h. The reaction mixture was cooled to room temperature and extracted with EtOAc. The remaining aqueous layer was extracted with CH2Cl2, and dried over Na2SO4. Concentration yielded 51.1 mg (24% yield) of the title compound as a white solid. 1H NMR (300 MHz, CDCl3) δ: 10.52 (bs, 1H), 7.40 (d, 1H, J=10.61), 7.39 (s, 1H), 7.11 (dd, 1H, J=10.24, J=1.83), 4.60 (s, 2H), 4.23 (s, 2H), 3.64 (m, 6H), 3.01 (s, 3H), 2.63 (t, 2H, J=6.22), 2.54, (m, 4H), 2.05 (s, 3H). MS (M+H) calcd for C19H23N3O4Cl2: 427.1; found: 428.11.
3-[(3,4-Dichloro-benzyl-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 12-B) was treated with paraformaldehyde and cyclopropylamine as described in the preparation of Compound 12. The title compound was extracted with EtOAc and the organic layer was washed with H2O, and dried over Na2SO4. After concentration, the resulting residue was triturated with EtOAc/Hexane (1:1) to give a white solid (52.7 mg, 30% yield). Mp=163–164° C. 1H NMR (300 MHz, CDCl3) δ: 9.97 (bs, 1H), 7.42, (d, 1H, J=8.42), 7.35 (s, 1H), 7.10 (dd, 1H, J=8.42, J=1.47), 4.60 (s, 2H), 4.09 (s, 2H), 3.02 (s, 3H), 2.81 (m, 1H), 0.86 (m, 4H). HRMS (M+H) calcd for C16H17N2Cl2O3: 355.0616. found: 355.0618.
3-[(3,4-Dichloro-benzyl-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 12-B) was treated with paraformaldehyde and methyl-ethane-1,2-diamine as described in the preparation of Compound 12. The title compound was triturated with EtOAc to give a white solid (7.8 mg, 4% yield). 1H NMR (300 MHz, CDCl3) δ: 9.14, (bs, 1H), 7.40, (d, 1H, J=8.05), 7.34 (s, 1H), 7.11 (dd, 1H, J=8.05, J=1.10), 4.58, (s, 2H), 4.34, (s, 2H), 3.94 (m, 2H), 3.25 (m, 2H), 3.03 (s, 3H), 2.78 (s, 3H). HRMS (M+H) calcd for C16H19N3Cl2O3: 372.0881. found: 372.0886.
3-[(3,4-Dichloro-benzyl-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 12-B) was treated with paraformaldehyde and 2,2-dimethyl-propylamine as described in the preparation of Compound 12. The resulting residue was purified by chromatography (YMC Combiprep ODS-A, 30 mm×50 mm, MeOH/H2O/0.1% TFA) to yield the title compound as a white powder (7.9 mg, 4% yield). 1H NMR (500 MHz, DMSO) δ: 7.42 (d, 1H, J=8.24), 7.36 (s, 1H), 7.12 (d, 1H, J=8.24), 4.61 (s, 2H), 4.26 (s, 2H), 3.27 (s, 2H), 3.04 (s, 3H), 0.93 (s, 9H). HRMS (M+H) calcd for C18H23N2Cl2O3: 385.1085; found: 385.1079.
3-[(3,4-Dichloro-benzyl-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 12-B) was treated with paraformaldehyde and 4-(2-amino-ethyl)benzenesulfonamide as described in the preparation of Compound 12. The resulting residue was purified by chromatography (YMC Combiprep ODS-A, 30 mm×50 mm, MeOH/H2O/0.1% TFA) to yield the title compound as a white powder (47.9 mg, 19% yield). Decomposition point=110–113° C. 1H NMR (500 MHz, DMSO) δ: 10.97 (s, 1H), 7.72, (m, 2H), 7.61 (d, 1H, J=8.54), 7.51 (s, 1H), 7.42 (m, 2H), 7.29 (m, 2H), 7.25, (m, 1H), 4.56, (s, 2H), 4.04, (s, 2H), 3.67 (m, 2H), 3.34 (m, 2H), 2.95 (d, 3H, J=6.41). HRMS (M−H) calcd for C21H20SN3Cl2O5: 496.0500. found: 496.0503.
3-[(3,4-Dichloro-benzyl-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 12-B) was treated with paraformaldehyde and β-alanine as described in the preparation of Compound 12. The resulting residue was purified by chromatography (YMC Combiprep ODS-A, 30 mm×50 mm, MeOH/H2O/0.1% TFA) to yield the title compound as an amber solid (48.5 mg, 25% yield). Mp=183–184° C. 1H NMR (500 MHz, DMSO) δ: 12.34 (s, 2H), 10.79 (s, 1H), 7.61 (d, 1H, J=8.54), 7.51 (s, 1H), 7.24 (bs, 1H), 4.57 (s, 2H), 4.06 (s, 2H), 3.58 (t, 2H, J=6.40), 2.96 (bs, 3H), 2.54 (t, 2H, J=7.02). HRMS (M−H) calcd for C16H15N2Cl2O5: 385.0358. found: 385.0364.
3-[(3,4-Dichloro-benzyl-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 12-B) was treated with paraformaldehyde and 3,4 dichloro-benzylamine as described in the preparation of Compound 12. The title compound was isolated as a white foam (155 mg, 66% yield). 1H NMR (500 MHz, CDCl3) δ: 10.70 (bs, 1H), 7.42 (d, 1H, J=3.05), 7.40 (d, 1H, J=3.05), 7.34 (d, 2H, J=1.83), 7.09 (dd, 2H, J=8.24, J=1.83), 4.62 (s, 2H), 4.57 (s, 2H), 4.06 (s, 2H), 3.00 (s, 3H). HRMS (M−H) calcd for C20H15N2Cl4O3: 470.9836. found: 470.9832.
3-[(3,4-Dichloro-benzyl-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 12-B) was treated with paraformaldehyde and glycine as described in the preparation of Compound 12. The title compound was isolated as a white solid (80 mg, 43% yield). Mp=178–180° C. 1H NMR (300 MHz, DMSO) δ: 12.97 (bs, 1H), 11.07 (s, 1H), 7.62 (d, 1H, J=8.41), 7.53 (s, 1H), 7.25 (d, 1H, J=6.95), 4.59 (s, 2H), 4.15 (s, 2H), 4.10 (s, 2H), 2.98 (s, 3H). HRMS (M−H) calcd for C15H13N2Cl2O5: 371.0201. found: 371.0216.
3-[(3,4-Dichloro-benzyl-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 12-B) was treated with paraformaldehyde and 4-(amino-methyl)pyridine as described in the preparation of Compound 12. The resulting residue was purified by chromatography (YMC Combiprep ODS-A, 30 mm×50 mm, MeOH/H2O/0.1% TFA) to yield the title compound as a sticky orange solid (10.5 mg, 5% yield). 1H NMR (300 MHz, DMSO) δ: 8.75 (d, 2H, J=6.22), 7.63–7.52 (m, 4H), 7.25 (d, 2H, J=7.31), 4.80 (s, 2H), 4.59 (s, 2H), 4.1 (s, 2H), 2.99 (bs, 3H). HRMS (M−H) calcd for C19H16N3Cl2O3: 404.0568. found: 404.0554.
3-[(3,4-Dichloro-benzyl-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 12-B) was treated with paraformaldehyde and 2-(aminomethyl)-1-ethylpyrrolidine as described in the preparation of Compound 12. The title compound was isolated as an orange solid (18.8 mg, 9% yield). 1H NMR (300 MHz, CDCl3) δ: 12.26 (bs, 1H), 7.43 (d, 1H, J=8.42), 7.36 (s, 1H), 7.13 (dd, 1H, J=8.05, J=1.46), 4.61 (s, 2H), 4.12 (s, 2H), 3.93 (m, 2H), 3.22 (t, 1H, J=8.78), 3.09 (s, 3H), 2.93 (m, 2H), 2.21–1.72 (m, 6H), 1.42 (t, 3H, J=6.95). HRMS (M−H) calcd for C20H24N3Cl2O3: 424.11948. found: 424.1200.
3-[(3,4-Dichloro-benzyl-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 12-B) was treated with paraformaldehyde and (2-aminoethyl)phosphonic acid diethylester as described in the preparation of Compound 12. Trituration with CH2Cl2/hexane gave the title compound (106 mg, 44% yield). 1H NMR (300 MHz, CDCl3) δ: 7.41 (d, 1H, J=8.05), 7.35 (s, 1H), 7.10 (dd, 1H, J=8.05, J=1.47), 4.60 (s, 2H), 4.25 (s, 2H), 4.11 (m, 4H), 3.77 (m, 2H), 3.02 (s, 3H), 2.17 (m, 2H), 1.31 (t, 6H, J=7.32). HRMS (M−H) calcd for C19H24PN2Cl2O6: 477.0749. found: 477.0749.
Compound 23-A (77 mg, 0.16 mmol) was stirred in acetic acid (4 mL) and concentrated HCl (1 mL) at 120° C. 18 hours. Mixture was cooled to room temp and concentrated. Residue was purified by chromatography (YMC Combiprep ODS-A, 30 mm×50 mm, MeOH/H2O/0.1% TFA) to give the title compound as a white powder (4.4 mg, 6.5% yield). 1H NMR (300 MHz, DMSO) δ: 7.61 (d, 1H, J=8.05), 7.52 (s, 1H), 7.24 (d, 1H, J=8.05), 4.58 (s, 2H), 4.09 (s, 2H), 3.55 (m, 2H), 2.96 (bs, 3H), 1.84 (m, 2H). HRMS (M−H) calcd for C15H16PN2Cl2O6: 421.0123. found: 421.0139.
3-[(3,4-Dichloro-benzyl-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 12-B) was treated with paraformaldehyde and -aminobutyric acid as described in the preparation of Compound 12. The resulting residue was purified by chromatography (YMC Combiprep ODS-A, 30 mm×50 mm, MeOH/H2O/0.1% TFA) to yield the title compound as a white powder (42.1 mg, 21% yield). 1H NMR (300 MHz, CDCl3) δ: 7.42 (d, 1H, J=8.05), 7.36 (s, 1H), 7.11 (dd, 1H, J=8.05, J=1.10), 4.60 (s, 2H), 4.21 (s, 2H), 3.58 (t, 2H, J=6.22), 3.03 (s, 3H), 2.40 (t, 2H, J=6.95), 1.96 (t, 2H, J=6.59). HRMS (M−H) calcd for C17H17N2Cl2O5: 399.0514. found: 399.0509.
3-[(3,4-Dichloro-benzyl-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 12-B) was treated with paraformaldehyde and asparagine as described in the preparation of Compound 12. The resulting residue was purified by chromatography (YMC Combiprep ODS-A, 30 mm×50 mm, MeOH/H2O/0.1% TFA) to yield the title compound as a white powder (12 mg, 6% yield). 1H NMR (300 MHz, DMSO) δ: 13.1 (bs, 1H), 11.05 (bs, 1H), 7.62 (d, 1H, J=8.42), 7.53 (s, 1H), 7.51 (s, 1H), 7.25 (bs, 1H), 6.97 (s, 1H), 4.90 (m, 1H), 4.58 (s, 2H), 4.07 (s, 2H), 2.96 (s, 3H), 2.73 (m, 2H). HRMS (M−H) calcd for C17H16N3Cl2O6: 428.0416. found: 428.0415.
3-[(3,4-Dichloro-benzyl-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 12-B) was treated with paraformaldehyde and 1-(2-aminoethyl)piperidine as described in the preparation of Compound 12. The resulting residue was purified by trituration with CH2Cl2 to yield the title compound as a white solid (81.2 mg, 38% yield). Decomposition point=178–182° C. 1H NMR (300 MHz, DMSO) δ: 10.99 (s, 1H), 7.62 (d, 1H, J=8.25), 7.52 (d, 1H, J=1.46), 7.26 (d, 1H, J=7.68), 4.60 (s, 2H), 4.13 (s, 2H), 3.81 (t, 2H, J=5.85), 3.48 (d, 2H, J=11.34), 3.27 (d, 2H. J=4.76), 3.00 (s, 2H), 2.86 (m, 3H), 1.77 (m, 6H). HRMS (M+H) calcd for C20H26N3Cl2O3: 426.1351. found: 426.1346.
3-[(3,4-Dichloro-benzyl-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 12-B) was treated with paraformaldehyde and N-acetylethylene diamine as described in the preparation of Compound 12. The resulting residue was purified by chromatography (YMC Combiprep ODS-A, 30 mm×50 mm, MeOH/H2O/0.1% TFA) to yield the title compound as a white powder (60.8 mg, 30% yield). 1H NMR (500 MHz, CDCl3) δ: 7.44 (d, 1H, J=8.24), 7.36 (s, 1H), 7.11 (dd, 1H, J=9.76, J=7.93), 6.77 (s, 1H), 4.60 (s, 2H), 4.29 (s, 2H), 3.68 (t, 2H, J=5.18), 3.55 (q, 2H, J=5.80), 3.05 (s, 3H), 2.02 (s, 3H). HRMS (M−H) calcd for C17H18N3Cl2O4: 398.0674. found: 398.0682.
3-[(3,4-Dichloro-benzyl-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 12-B) was treated with paraformaldehyde and 7-amino-heptanoic acid as described in the preparation of Compound 12. The resulting residue was purified by chromatography (YMC Combiprep ODS-A, 30 mm×50 mm, MeOH/H2O/0.1% TFA) to yield the title compound as a white powder (11.2 mg, 5% yield). 1H NMR (500 MHz, CDCl3) δ: 11.95 (bs, 1H), 11.00 (s, 1H), 7.61 d, 1H, J=8.24), 7.52 (s, 1H), 7.25 (s, 1H), 4.58 (s, 2H), 4.04 (s, 2H), 3.36 (m, 2H), 2.98 (s, 3H), 2.18 (t, 2H, J=7.33), 1.48 (m, 4H), 1.26 (m, 4H). HRMS (M+H) calcd for C20H25N2Cl2O5: 443.1140. found: 443.1153.
3-[(3,4-Dichloro-benzyl-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 12-B) was treated with paraformaldehyde and L-anserine nitrate as described in the preparation of Compound 12. The resulting residue was purified by chromatography (YMC Combiprep ODS-A, 30 mm×50 mm, MeOH/H2O/0.1% TFA) to yield the title compound as a white solid (3.3 mg, 1% yield). 1H NMR (500 MHz, CDCl3) δ: 8.56 (s, 1H), 7.36 (d, 1H, J=8.24) 7.29 (m, 1H), 7.20 (s, 1H), 7.04 (m, 1H), 4.69 (m, 1H), 4.52 (s, 2H), 4.05 (s, 2H), 3.76 (s, 3H), 3.65 (t, 2H, J=5.80), 3.14 (m, 2H), 2.94 (s, 2H), 2.51 (m, 2H). HRMS (M+H) calcd for C23H26N5Cl2O6: 538.1260. found: 538.1280.
3-[(3,4-Dichloro-benzyl-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 12-B) was treated with paraformaldehyde and N-propylamino imidazole as described in the preparation of Compound 12. The resulting residue was purified by chromatography (YMC Combiprep ODS-A, 30 mm×50 mm, MeOH/H2O/0.1% TFA) to yield the title compound as an amber foam (59.1 mg, 28% yield). 1H NMR (300 MHz, DMSO) δ: 14.47 (bs, 1H), 11.05 (bs, 1H), 9.10 (s, 1H), 7.82 (s, 1H), 7.70 (s, 1H), 7.62 (d, 1H, J=8.05), 7.52 (s, 1H), 7.25 (d, 1H, J=7.32), 4.59 (s, 2H), 4.19 (t, 1H, J=6.95), 4.09 (s, 2H), 3.41 (t, 2H, J=6.59), 2.98 (s, 3H), 2.13 (t, 2H, J=6.59). HRMS (M+H) calcd for C19H21N4Cl2O3: 423.0990. found: 423.0982.
3-[(3,4-Dichloro-benzyl-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 12-B) was treated with paraformaldehyde and methylamine thiophene as described in the preparation of Compound 12. The resulting residue was purified by chromatography (YMC Combiprep ODS-A, 30 mm×50 mm, MeOH/H2O/0.1% TFA) to yield the title compound as a white powder (58.7 mg, 28% yield). 1H NMR (300 MHz, CDCl3) δ: 7.41 (d, 1H, J=8.05), 7.33 (d, 1H, J=1.83), 7.26 (m, 1H), 7.08 (dd, 1H, J=8.05, J=1.83), 6.97 (m, 2H), 4.85 (s, 2H), 4.57 (s, 2H), 4.11 (s, 2H), 3.00 (s, 3H). HRMS (M−H) calcd for C18H15SN2Cl2O3: 409.0180. found: 409.0190.
A mixture of 1-methyl piperazine (0.55 mL, 5.0 mmol), potassium carbonate (3.49 g, 25 mmol) and chloroacetonitrile (0.34 mL, 5.6 mmol) in acetonitrile (3 mL) was stirred at room temperature 8 h. The mixture was diluted with diethyl ether and filtered and concentrated to yield the title compound as yellow crystals (536.6 mg, 77% yield). 1H NMR (500 MHz, CDCl3) δ: 3.50 (s, 2H), 2.66 (t, 4H, J=4.58), 2.52 (bs, 4H), 2.33 (s, 3H).
To a suspension of lithium aluminum powder (0.16 g, 4.2 mmol) and diethyl ether (5.0 mL) cooled to 0° C., was added dropwise a solution of the above compound (0.536 g, 3.86 mmol) dissolved in diethyl ether/THF (14 mL, 1:1). The resulting mixture was stirred at room temperature for 5 h. The mixture was cooled to 0° C. and 2N aqueous NaOH added dropwise. The mixture was stirred for 20 min and the precipitate was filtered. The organic solution was concentrated and the resulting oil was dissolved in ethylacetate. A solution of 4N HCl in dioxane (0.97 mL) added and white precipitate filtered to yield the title compound as the HCl salt. 1H NMR (500 MHz, DMSO) δ: 9.48 (s, 2H), 4.18 (m, 6H), 3.75 (m, 2H), 3.68 (m, 2H), 3.26 (m, 2H), 2.82 (s, 3H).
3-[(3,4-Dichloro-benzyl-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 12-B) was treated with paraformaldehyde and 2-(4-methyl-piperazine-1-yl)-ethyl amine as described in the preparation of Compound 12. The resulting residue was purified by chromatography (YMC Combiprep ODS-A, 30 mm×50 mm, MeOH/H2O/0.1% TFA) to yield the title compound as an amber glass (27 mg, 12% yield). 1H NMR (300 MHz, DMSO) δ: 7.63 (d, 1H, J=8.42), 7.52 (s, 1H), 7.25 (d, 1H, J=7.31), 4.59 (s, 2H), 4.24 (m, 10H), 4.11 (s, 2H), 3.58 (s, 2H), 2.98 (s, 3H), 2.78 (s, 3H). HRMS (M−H) calcd for C20H25N4Cl2O3: 439.1303. found: 439.1311.
3-[(3,4-Dichloro-benzyl-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 12-B) was treated with paraformaldehyde and 4-aminobutyrate hydrochloride as described in the preparation of Compound 12. The resulting residue was purified by chromatography (YMC Combiprep ODS-A, 30 mm×50 mm, MeOH/H2O/0.1% TFA) to yield the title compound as an amber oil (18.9 mg, 9% yield). 1H NMR (300 MHz, CDCl3) δ: 7.41 (m, 2H), 7.09 (m, 1H), 4.60 (s, 2H), 4.18 (s, 2H), 3.65 (s, 3H), 3.55 (t, 2H, J=7.32), 3.03 (s, 3H), 2.36 (t, 2H, J=7.32), 1.94 (t, 2H, J=6.95). HRMS (M+H) calcd for C18H21N2Cl2O53: 415.0827. found: 415.0831.
3-[(3,4-Dichloro-benzyl-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 12-B) was treated with paraformaldehyde and β-alanine methyl ester as described in the preparation of Compound 12. The resulting residue was purified by chromatography (YMC Combiprep ODS-A, 30 mm×50 mm, MeOH/H2O/0.1% TFA) to yield the title compound as an amber oil (34.3 mg, 17% yield). 1H NMR (300 MHz, CDCl3) δ: 7.41 (m, 2H), 7.09 (m, 1H), 4.59 (s, 2H), 4.27 (s, 2H), 3.77 (m, 2H), 3.67 (s, 3H), 3.03 (s, 3H), 2.69 (t, 2H, J=6.22). HRMS (M+H) calcd for C17H19N2Cl2O5: 401.0671. found: 401.0669.
3-[(3,4-Dichloro-benzyl-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 12-B) was treated with paraformaldehyde and H-gly-gly-β-ala-gly-OH as described in the preparation of Compound 12. The reaction mixture was diluted with water and ethylacetate. The white solids were filtered as the title compound (88.2 mg, 32% yield). 1H NMR (300 MHz, DMSO) δ: 8.31 (t, 1H, J=5.49), 8.20 (t, 1H, J=5.49), 7.92 (t, 1H, J=5.49), 7.61 (d, 1H, J=8.42), 7.52 (s, 1H), 7.25 (d, 1H, J=6.59), 4.59 (s, 2H), 4.09 (s, 4H), 3.69 (m, 6H), 2.94 (bs, 3H), 2.30(t, 2H, J=7.31). HRMS (M−H) calcd for C22H24N5Cl2O8: 556.1002. found: 556.0994.
3-[(3,4-Dichloro-benzyl-methyl-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 12-B) was treated with paraformaldehyde and 4-(aminomethyl)benzoic acid as described in the preparation of Compound 12. The resulting residue was triturated with chloroform to give the title compound as a white solid (160 mg, 71% yield). 1H NMR (300 MHz, DMSO) δ: 12.94 (s, 1H), 11.14 (s, 1H), 8.32 (s, 1H), 7.93 (d, 2H, J=8.05), 7.60 (d, 1H, J=8.45), 7.50 (s, 1H), 7.35 (d, 2H, J=7.84), 7.23 (ds, 1H, 1=8.05), 4.66 (s, 2H), 4.57 (s, 2H), 3.99 (s, 2H), 2.97 (s, 3H). HRMS (M+H) calcd for C21H17N2Cl2O5: 447.0515. found: 447.0530.
To a suspension of (2,2-Dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetic acid, Compound III-A, (4.5 g, 26.2 mmol) in benzene (30 mL) was added oxalyl chloride (15 mL) and the resulting mixture was heated at reflux for 1 h. The mixture was cooled to room temp. and concentrated. The residue was dissolved in methylene chloride (30 mL) and cooled to 0° C. To this was added a suspension of (3,4-dichloro-benzyl)-methyl-amine hydrochloride salt (5.0 g, 22.1 mmol) in methylene chloride (30 mL) and pyridine (18 mL). The resulting mixture was stirred at room temp for 18 h then diluted with 1N HCl. The aqueous phase was saturated with sodium chloride and extracted with methylene chloride (3 times). The organic layers were combined, dried (Na2SO4), and concentrated. The brown oil was purified over silica gel eluting with ethyl acetate/hexane (1:1) to give the title compound as a pale yellow oil that solidified to a white solid upon standing (6.2 g, 82% yield). 1H NMR (300 MHz, DMSO) δ: 7.44–7.10 (m, 3H), 6.16 (s, 0.66H), 6.08 (s, 0.33H), 4.59 (s, 1.33H), 4.53 (s, 0.67H), 3.02 (s, 2H), 2.97 (s, 1H), 1.74 (s, 4H), 1.69 (s, 2H).
A mixture of paraformaldehyde (0.015 g, 0.5 mmol) and methanol (1.5 mL) was warmed to 55° C. Added to this mixture was H-β-Ala-β-ala-OH (0.08 g, 0.5 mmol) and the solution was stirred 5 min. N-(3,4-Dichloro-benzyl)-2-(2,2-dimethyl-5-oxo[1,3]dioxolane-4-ylidene)-N-methyl-acetamide (0.1715 g, 0.5 mmol) was added and the mixture was stirred at 55° C. for 45 min. The mixture was concentrated and the resulting residue was triturated with ethylacetate and filtered to give the desired product as a white solid (0.0358 g, 16% yield). 1H NMR (300 MHz, DMSO) δ: 2.35 (t, J=6.7 Hz, 4H), 2.96 (s, 3H), 3.32 (m, 2H), 3.57 (m, 2H), 4.02 (s, 2H), 4.57 (s, 2H), 7.23 (d, J=6.6 Hz, 1H), 7.51 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 8.05 (t, J=10.3 Hz, 1H), 10.97 (s, 1H), 12.19 (s, 1H). HRMS (M+H) calcd for C19H21N3Cl2O6: 456.07292. found: 456.0738.
N-(3,4-Dichloro-benzyl)-2-(2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-methyl-acetamide (Compound 37-A) was treated with paraformaldehyde and N-(2-amino-ethyl)-3-morpholin-4-yl-propionamide as described in the preparation of Compound 37. The desired product was isolated as a white solid (0.0134 g, 5% yield). 1H NMR (300 MHz, CDCl3) δ: 2.41 (m, 2H), 2.64 (bs, 2H), 2.72 (bs, 2H), 2.98 (m, 2H), 3.03 (s, 3H), 3.51 (t, J=5.6 Hz, 2H), 3.63 (t, J=5.2 Hz, 2H), 3.78 (m, 4H), 4.35 (s, 2H), 4.59 (s, 2H), 7.11 (d, J=8.1 Hz, 1H), 7.35–7.44 (m, 3H), 8.27 (t, 1H). HRMS (M+H) calcd for C22H28N4Cl2O5: 497.13585. found: 497.1377.
N-(3,4-Dichloro-benzyl)-2-(2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-methyl-acetamide (Compound 37-A) was treated with paraformaldehyde and H-β-ala-gly-OH as described in the preparation of Compound 37. The title compound was purified by chromatography (YMC Combiprep ODS-A, 30 mm×50 mm, MeOH/H2O/0.1% TFA) to yield the title compound as an amber powder (0.0158 g, 7% yield). 1H NMR (300 MHz, DMSO) δ: 2.45 (t, J=6.9 Hz, 2H), 2.98 (m, 3H), 3.60 (m, 2H), 3.73 (d, J=5.7 Hz, 2H), 4.06 (s, 2H), 4.57 (s, 2H), 7.23 (d, J=7.2, 1H), 7.65 (d, J=8.4 Hz, 1H), 8.32 (m, 1H), 10.98 (bs, 1H), 12.50 (bs, 1H). HRMS (M−H) calcd for C18H19N3Cl2O6: 442.05727. found: 442.0584.
N-(3,4-Dichloro-benzyl)-2-(2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-methyl-acetamide (Compound 37-A) was treated with paraformaldehyde and β-alaninamide as described in the preparation of Compound 37. The title compound was purified by chromatography (YMC Combiprep ODS-A, 30 mm×50 mm, MeOH/H2O/0.1% TFA) to yield the title compound as an amber powder (0.0158 g, 7% yield). 1H NMR (300 MHz, CDCl3) δ: 2.68 (t, J=5.9 Hz, 2H), 3.02 (s, 3H), 3.79 (d, J=5.9 Hz, 2H), 4.29 (s, 2H), 4.58 (s, 2H), 6.23 (s, 1H), 6.59 (s, 1H), 7.09 (dd, J=8.4 Hz, J=10.3 Hz, 1H), 7.34 (d, J=1.8 Hz, 1H), 7.42 (d, J=8.4 Hz, 1H). HRMS (M−H) calcd for C16H17N3Cl2O4: 384.05179. found: 384.0517.
N-(3,4-Dichloro-benzyl)-2-(2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-methyl-acetamide (Compound 37-A) was treated with paraformaldehyde and 4-(aminomethyl)-benzoate hydrochloride salt with 1 equivalent of triethyl amine as described in the preparation of Compound 37. The title compound was purified by chromatography (YMC Combiprep ODS-A, 30 mm×50 mm, MeOH/H2O/0.1% TFA) to yield the title compound as a yellow foam (0.060 g, 26% yield). 1H NMR (300 MHz, CDCl3) δ: 2.98 (s, 3H), 3.91 (s, 3H), 4.05 (s, 2H), 4.56 (s, 2H), 7.07 (dd, J=1.8 Hz, J=8.1 Hz, 1H), 7.30 (m, 3H), 7.40 (m, 1H), 8.00 (d, J=8.1 Hz, 1H). HRMS (M−H) calcd for C22H20N2Cl2O5: 461.0671. found: 461.0690.
N-(3,4-Dichloro-benzyl)-2-(2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-methyl-acetamide (Compound 37-A) was treated with paraformaldehyde and 3-aminopropionitirile with 1 equivalent of triethyl amine as described in the preparation of Compound 37. The title compound was purified by chromatography (YMC Combiprep ODS-A, 30 mm×50 mm, MeOH/H2O/0.1% TFA) to yield the title compound as a yellow foam (0.0819 g, 45% yield). 1H NMR (300 MHz, CDCl3) δ: 2.73 (t, J=6.3 Hz, 2H), 3.04 (s, 3H), 3.78 (t, J=6.3 Hz, 2H), 4.40 (s, 2H), 4.60 (s, 2H), 7.11 (dd, J=1.8 Hz, J=8.1 Hz, 1H), 7.35 (d, J=1.5 Hz, 1H), 7.43 (d, J=8.4 Hz, 1H). HRMS (M−H) calcd for C16H15N3Cl2O3: 366.04122. found: 366.0428.
N-(3,4-Dichloro-benzyl)-2-(2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-methyl-acetamide (Compound 37-A) was treated with paraformaldehyde and N-(3-aminopropyl)morpholine as described in the preparation of Compound 37. The title compound was isolated as a white solid (0.1345 g, 61% yield). 1H NMR (500 MHz, DMSO) δ: 1.69 (t, J=6.4 Hz, 2H), 2.25 (s, 2H), 2.32 (s, 4H), 2.97 (s, 3H), 3.40 (t, J=6.4 Hz, 2H), 3.55 (s, 4H), 4.05 (s, 2H), 4.59 (s, 2H), 7.24 (s, 1H), 7.51 (s, 1H), 7.62 (d, J=8.3 Hz, 1H). HRMS (M−H) calcd for C20H25N3Cl2O4: 442.13005. found: 442.1296.
A solution of methoxylamine hydrochloride (13.4 g, 0.16 mol) in a mixture of water (150 ml) and tetrahydrofuran (50 ml) was treated with sodium acetate (11.2 g, 0.136 mol) followed by 4-fluorobenzaldehyde (11.57 g, 93.2 mmol) and the resulting mixture was stirred at 22° C. for 4 hours. The reaction mixture was then diluted with ether, washed with brine and dried over anhydrous magnesium sulfate. Evaporation of the solvent under reduced pressure gave 14.3 g of the crude title material as a clear oil which was used as such for the next step. Distillation of an aliquot in vacuo gave a clear oil; bp 45–50° C./0.5 torr. 1HNMR 400 MHz (CDCl3) δ (ppm): 3.99 (3H, s), 7.09 (2H, m), 7.6 (2H, m), 8.06 (1H, s).
A solution of 4-fluorobenzaldehyde-O-methyloxime (93.2 mmol) in dichloromethane (150 ml) was treated with sodium cyanoborohydride (9.18 g, 0.146 mol) followed by 120 ml of 2 N hydrochloric acid in methanol added dropwise over 30 minutes. After 96 h at 22° C., the solvent was evaporated under reduced pressure and the residue was slurried with water and the pH was adjusted to 9 with 2 N aqueous sodium hydroxide. The aqueous phase was extracted twice with dichloromethane and the combined organic extracts were washed with brine, dried (magnesium sulfate) and concentrated under reduced pressure. The residual oil was chromatographed on silica gel (elution toluene-ethyl acetate 0–10% yield) and gave 5.92 g (41% yield) of the title amine as a clear oil. 1HNMR 400 MHz (CDCl3) δ (ppm): 3.49 (3H, s), 4.01 (2H, s), 5.69 (1H, broad s), 7.01 (2H, m), 7.31 (2H, m). The hydrochloride salt was obtained as a white solid: mp 170–171° C. Anal. calcd for C8H10FNO—HCl: C, 50.14; H, 5.78; N, 7.31. Found: C, 50.31; H, 5.80; N, 7.26.
In an alternative procedure a solution of 4-fluorobenzaldehyde O-methyloxime (0.82 g, 5.35 mmol) in acetic acid (8 ml) was treated at 10° C. with sodium cyanoborohydride (0.67 g, 10.7 mmol) added in small portions over 10 min and the resulting solution was stirred at 25° C. for 18 h. The solvent was evaporated under reduce pressure (co-evaporation with toluene twice) and the residue was slurried with water and the pH was adjusted to 9 with 2 N aqueous sodium hydroxide. The aqueous phase was extracted twice with ether and the combined organic extracts were washed with brine, dried (magnesium sulfate) and concentrated under reduced pressure. The residual oil was chromatographed on silica gel (elution hexane-ethyl acetate, 8:2) and distilled in vacuo to give 0.62 g (75% yield) of the title amine as a clear oil.
A solution of (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl chloride (2.45 g, 12.9 mmol) in dichloromethane (15 ml) was added dropwise over 10 minutes to a cold (0–5° C.) mixture of N-4-fluorobenzyl-O-methyl-hydroxylamine (2.0 g, 12.9 mmol) and pyridine (2.1 ml, 25.7 mmol) in dichloromethane (50 ml). The cooling bath was then removed and the solution was stirred at 22° C. for 30 minutes. The reaction mixture was then quenched by the addition of water and ethyl acetate. The organic phase was washed successively with 0.1 N hydrochloric acid, saturated sodium bicarbonate, brine and dried (magnesium sulfate). Evaporation of the solvent and chromatography of the residue on silica gel (toluene-ethyl acetate, 8:2) gave 3.72 g (93% yield) of the title amide as white crystals: mp 111° C. (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.75 (6H, s), 3.68 (3H, s), 4.79 (2H, s), 6.38 (1H, s), 7.0 (2H, m), 7.34 (2H, m). Anal. calcd for C15H16FNO5: C, 58.25; H, 5.21; N, 4.52. Found: C, 58.33; H, 5.38; N, 4.51.
A solution of 2-(2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-(4-fluorobenzyl)-N-methoxy-acetamide (0.64 g, 2.07 mmol) in methanol (50 ml), was treated at 22° C. with 0.025 ml of a 4.6 M solution of sodium methoxide in methanol and the resulting mixture was stirred for 2 h at the same temperature. The reaction mixture was then quenched by the addition of 1 ml of 1 N hydrochloric acid and the solvent was concentrated under reduced pressure. The residue was diluted with ethyl acetate, washed with water and brine and then dried over anhydrous magnesium sulfate. Evaporation of the solvent and crystallization of the residue from a mixture of ethyl acetate and hexane gave 0.559 g (95% yield) of the title ester as white prisms; mp 71° C. 1HNMR 400 MHz (CDCl3) δ (ppm), mixture of rotamers, major: 3.69 (3H, s, OCH3), 3.89 (3H, s, OCH3), 4.80 (2H, s, NCH2), 6.45 (1H, s, CH), 7.03 (2H, m, aromatics), 7.30 (2H, m, aromatics). Anal. calcd for C13H14FNO5: C, 55.12; H, 4.98; N, 4.94. Found: C, 54.95; H, 4.73; N, 4.67.
Method 44A: 3-[(4-Fluorobenzyl)-methoxy-carbamoyl]-2-hydroxy-acrylic acid methyl ester was treated with paraformaldehyde and methylamine as described in the preparation of Compound 12 to give the title compound as a white solid (48% yield); mp 125° C., dec. 1HNMR 400 MHz (CDCl3) δ (ppm): 3.1 (3H, s, NCH3), 3.72 (3H, s, OCH3), 4.15 (2H, s, NCH2), 4.84 (2H, s, NCH2), 7.04 (2H, m, aromatics), 7.32 (2H, m, aromatics). Anal. calcd for C14H15FN2O4: C, 57.14; H, 5.13; N, 9.52. Found: C, 56.87; H, 5.12; N, 9.42.
Method 44B: A solution of 2 M methylamine in tetrahydrofuran (0.58 ml, 1.16 mmol) was added to a mixture of paraformaldehyde (0.35 g, 1.16 mmol, equivalent of formaldehyde) in methanol (3 ml) and the resulting mixture was heated at 60° C. for 5 min. Then solid 2-(2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-(4-fluorobenzyl)-N-methoxy-acetamide (0.30 g, 0.97 mmol) was added all at once and the resulting mixture was maintained at 60° C. for another 60 min. The reaction mixture was then quenched by the addition of ethyl acetate and pH 2 phosphate buffer. The organic phase was washed with brine, dried (anhydrous magnesium sulfate) and concentrated under reduced pressure. Recrystallization of the solid residue from a mixture of ethyl acetate and hexane gave 0.181 g (64% yield) of the title material as a white solid.
3-[(4-Fluoro-benzyl)-methoxy-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 44-D) was treated with paraformaldehyde and N-(2-aminoethyl)morphiline as described in the preparation of Compound 13. The reaction mixture was diluted with water and extracted the CH2Cl2. After concentration of the organic phase, the residue was triturated with EtOAc and the solid filtered to yield the title compound as a white solid (0.1444 g, 37% yield). 1H NMR (500 MHz, DMSO) δ: 3.08 (d, J=11.1 Hz, 2H), 3.41 (d, J=5.2 Hz, 2H), 3.51 (d, J=12.2 Hz, 2H), 3.69 (t, J=11.9 Hz, 2H), 3.75 (s, 3H), 3.81 (s, 2H), 3.98 (d, 1=13.2 Hz, 2H), 4.25 (s, 2H), 4.88 (s, 2H), 7.18 (t, J=8.8 Hz, 2H), 7.37 (m, 2H), 10.11 (s, 1H), 11.42 (s, 1H). HRMS (M+H) calcd for C19H24FN3O5: 392.16217. found: 392.1627.
3-[(4-Fluoro-benzyl)-methoxy-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 44-D) was treated with paraformaldehyde and N-(2-aminoethyl)piperazine as described in the preparation of Compound 13. The mixture was purified by chromatography (YMC Combiprep ODS-A, 30 mm×50 mm, MeOH/H2O/0.1% TFA) to yield the title compound as a white powder (0.0110 g, 4.3% yield). 1H NMR (500 MHz, CDCl3) δ: 2.55 (t, J=6.2 Hz, 2H), 2.71 (s, 4H), 2.98 (s, 4H), 3.47 (t, J=5.5 Hz, 2H), 3.57 (s, 3H), 4.03 (s, 2H), 4.68 (s, 2H), 6.87 (s, 2H), 7.17 (m, 2H), 9.72 (bs, 1H), 11.45 (bs, 1H). HRMS (M+H) calcd for C19H25FN4O4: 391.17816. found: 391.1786.
Reaction of 3,4-dichlorobenzaldehyde with methoxylamine hydrochloride followed by reduction with sodium cyanoborohydride as described in the preparation of Compound 44-A and 44-B gave the title hydroxylamine as a clear oil. 1HNMR 400 MHz (CDCl3) δ (ppm): 3.48 (3H, s), 3.99 (2H, s), 5.74 (1H, broad s), 7.20 (1H, dd, J=2.0 Hz and J=8.1 Hz), 7.40 (1H, d, J=8.1 Hz), 7.47 (1H, d, J=2.0 Hz).
Reaction of (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl chloride with N-3,4-dichlorobenzyl-O-methyl-hydroxylamine as described in the preparation of Compound 44-C gave the title amide as a white solid (94% yield): mp 119–120° C. (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.76 (6H, s), 3.71 (3H, s), 4.72 (2H, s), 6.38 (1H, s), 7.20 (1H, dd, J=2.0 Hz and J=8.5 Hz), 7.40 (1H, d, J=8.5 Hz), 7.46 (1H, d, J=2.0 Hz). Anal. calcd for C15H15Cl2NO5: C, 50.02; H, 4.20; N, 3.89. Found: C, 50.12; H, 4.12; N, 3.80.
N-(3,4-Dichlorobenzyl)-2-(2,2-dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-methoxy-acetamide was treated with methanol as described in the preparation of Compound 44-D and gave the title ester as white crystals (95% yield); mp 111° C. 1HNMR 400 MHz (CDCl3) δ (ppm): 3.72 (3H, s, OCH3), 3.90 (3H, s, OCH3), 4.77 (2H, s, NCH2), 6.46 (1H, s, CH), 7.17 (1H, dd, J=2.0 Hz, J=8.0 Hz, aromatic), 7.42 (1H, d, J=8.0 Hz, aromatic), 7.43 (1H, d, J=2.0 Hz, aromatic). Anal. calcd for C13H13Cl2NO5: C, 46.73; H, 3.92; N, 4.19. Found: C, 46.95; H, 3.82; N, 3.97.
3-[(3,4-Dichlorobenzyl)-methoxy-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 47-C) was treated with paraformaldehyde and methylamine as described in the preparation of Compound 12, yielding the title compound as a white solid (49% yield); mp 149° C. 1HNMR 400 MHz (CDCl3) δ (ppm): 3.11 (3H, s, NCH3), 3.75 (3H, s, OCH3), 4.15 (2H, s, NCH2), 4.82 (2H, s, NCH2), 7.18 (1H, dd, J=2.0 Hz, J=8.2 Hz, aromatic), 7.42 (1H, d, J=8.2 Hz, aromatic), 7.43 (1H, d, J=2.0 Hz, aromatic). Anal. calcd for C14H14Cl2N2O4: C, 48.72; H, 4.09; N, 8.12. Found: C, 48.81; H, 4.04; N, 7.99.
Reaction of 3-chloro-4-fluorobenzaldehyde with methoxylamine hydrochloride followed by reduction with sodium cyanoborohydride as described in the preparation of Compound 44-A and 44-B gave the title hydroxylamine as a clear oil. 1HNMR 400 MHz (CDCl3) δ (ppm): 3.48 (3H, s), 3.98 (2H, s), 5.72 (1H, broad s), 7.10 (1H, t), 7.22 (1H, m), 7.42 (1H, m).
Reaction of (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl chloride with N-(3-chloro-4-fluorobenzyl)-O-methyl-hydroxylamine as described in the preparation of Compound 44-C gave the title amide as a white solid (91% yield): mp 110–111° C. (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.76 (6H, s), 3.71 (3H, s), 4.75 (2H, s), 6.38 (1H, s), 7.09 (1H, t, J=8.8 Hz), 7.23 (1H, m), 7.41 (1H, dd, J=2.4 Hz and J=6.8 Hz). Anal. calcd for C15H15ClFNO5: C, 52.41; H, 4.39; N, 4.07. Found: C, 52.25, H, 4.36, N, 3.87.
N-(3-Chloro-4-fluorobenzyl)-2-(2,2-dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-methoxy-acetamide was treated with methanol as described in the preparation of Compound 44-D and gave the title ester as white crystals (54% yield); mp 97–98° C. 1HNMR 400 MHz (CDCl3) δ (ppm): 3.72 (3H, s, OCH3), 3.90 (3H, s, OCH3), 4.77 (2H, s, NCH2), 6.45 (1H, s, CH), 7.11 (1H, m, aromatic), 7.2 (1H, m, aromatic), 7.38 (1H, m, aromatic). Anal. calcd for C13H13ClFNO5: C, 49.14; H, 4.12; N, 4.40. Found: C, 48.95; H, 3.96; N, 4.16.
3-[(3-Chloro-4-fluorobenzyl)-methoxy-carbamoyl]-2-hydroxy-acrylic acid methyl ester (48-C) was treated with paraformaldehyde and methylamine as described in the preparation of Compound 12 to give the title compound as a white solid (60% yield); mp 148° C. dec. 1HNMR 400 MHz (CDCl3) δ (ppm): 3.10 (3H, s, NCH3), 3.75 (3H, s, OCH3), 4.15 (2H, s, NCH2), 4.81 (2H, s, NCH2), 7.12 (1H, m, aromatic), 7.22 (1H, m, aromatic), 7.38 (1H, m, aromatic). Anal. calcd for C14H14Cl2N2O4: C, 51.15; H, 4.29; N, 8.52. Found: C, 51.19; H, 4.17; N, 8.50.
Reduction 3-fluorobenzaldehyde O-methyloxime with sodium cyanoborohydride as described in the preparation of Compounds 44-A and 44-B gave the title hydroxylamine as a clear oil (60% yield). 1HNMR 400 MHz (CDCl3) δ (ppm): 3.50 (3H, s, OCH3), 4.04 (2H, s, NCH2), 5.75 (1H, broad s, NH), 6.95–7.32 (4H, m, aromatics). The hydrochloride salt was obtained as a white solid: mp 130–131° C. (dec.). Anal. calcd for C8H10FNO—HCl: C, 50.14; H, 5.78; N, 7.31. Found: C, 50.10; H, 5.73; N, 7.38.
Reaction of (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl chloride with N-(3-fluorobenzyl)-O-methyl-hydroxylamine as described in the preparation of Compound 44-C and gave the title amide as a white solid (94% yield): mp 110–111° C. (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.76 (6H, s, CH3), 3.70 (3H, s, OCH3), 4.82 (2H, s, NCH2), 6.40 (1H, s, CH), 6.96–7.32 (4H, m, aromatics). Anal. calcd. for C15H16FNO5: C, 58.25; H, 5.21; N, 4.52. Found: C, 58.00; H, 5.30; N, 4.49.
N-(3-Fluorobenzyl)-2-(2,2-dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-methoxy-acetamide was treated with methanol as described in the preparation of Compound 44-D and gave the title ester as white crystals (53% yield); mp 73–75° C. 1HNMR 400 MHz (CDCl3) δ (ppm): 3.71 (3H, s, OCH3), 3.90 (3H, s, OCH3), 4.82 (2H, s, NCH2), 6.47 (1H, s, CH), 6.98–7.34 (4H, m, aromatics). Anal. calcd for C13H14FNO5: C, 55.12; H, 4.98; N, 4.94. Found: C, 55.18; H, 5.04; N, 5.02.
3-[(3-Fluorobenzyl)-methoxy-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 49-C) was reacted with paraformaldehyde and methylamine as described in the preparation of Compound 12 to give the title compound as a white solid (60% yield); mp 119° C. dec. 1HNMR 400 MHz (CDCl3) δ (ppm): 3.11 (3H, s, NCH3), 3.74 (3H, s, OCH3), 4.17 (2H, s, NCH2), 4.87 (2H, s, NCH2), 6.99–7.35 (4H, m, aromatics). Anal. calcd for C14H15FN2O4: C, 57.14; H, 5.14; N, 9.52. Found: C, 57.04; H, 5.02; N, 9.42.
Reaction of 2-fluorobenzaldehyde with methoxylamine hydrochloride as described in the preparation of Compound 44-A gave the title oxime ether as a clear oil (98% yield). HPLC indicated a 91:9 mixture of E- to Z-isomers. 1HNMR 400 MHz (CDCl3) δ (ppm): (E-isomer) 3.99 (3H, s, OCH3), 7.07 (1H, m, aromatic), 7.14 (1H, m, aromatic), 7.34 (1H, m, aromatic), 7.82 (1H, m, aromatic), 8.31 (1H, s, CH).
Reduction of 2-fluorobenzaldehyde O-methyloxime with sodium cyanoborohydride as described in the preparation of Compound 44-B gave the title hydroxylamine as a clear oil (74% yield). 1HNMR 400 MHz (CDCl3) δ (ppm): 3.52 (3H, s, OCH3), 4.11 (2H, s, NCH2), 5.78 (1H, broad s, NH), 7.05 (1H, m, aromatic), 7.11 (1H, m, aromatic), 7.27 (1H, m, aromatic), 7.38 (1H, m, aromatic). The hydrochloride salt was obtained as a white solid: mp 138–143° C. (dec.). Anal. calcd. for C8H10FNO—HCl: C, 50.14; H, 5.78; N, 7.31. Found: C, 50.37; H, 5.71; N, 7.18.
Reaction of (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl chloride with N-(2-fluorobenzyl)-O-methyl-hydroxylamine as described in the preparation of Compound 44-C gave the title amide as a white solid (84% yield): mp 109–111° C. (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.75 (6H, s, CH3), 3.72 (3H, s, OCH3), 4.92 (2H, s, NCH2), 6.40 (1H, s, CH), 7.03–7.12 (2H, m, aromatics), 7.24–7.30 (1H, m, aromatic), 7.4 (1H, m, aromatic). Anal. calcd. for C15H16FNO5: C, 58.25; H, 5.21; N, 4.52. Found: C, 58.47; H, 5.16; N, 4.66.
N-(2-Fluorobenzyl)-2-(2,2-dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-methoxy-acetamide was treated with methanol as described in the preparation of Compound 44-D and gave the title ester as a white syrup (59% yield). 1HNMR 400 MHz (CDCl3) δ (ppm): 3.73 (3H, s, OCH3), 3.89 (3H, s, OCH3), 4.93 (2H, s, NCH2), 6.47 (1H, s, CH), 7.05–7.36 (4H, m, aromatics). Anal. calcd for C13H14FNO5: C, 55.12; H, 4.98; N, 4.94. Found: C, 54.91; H, 5.23; N, 4.86.
3-[(2-Fluorobenzyl)-methoxy-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 50-D) was treated with paraformaldehyde and methylamine as described in the preparation of Compound 12 to give the title compound as a white solid (58% yield); mp 147° C. dec. 1HNMR 400 MHz (CDCl3) δ (ppm): 3.1 (3H, s, NCH3), 3.76 (3H, s, OCH3), 4.15 (2H, s, NCH2), 4.97 (2H, s, NCH2), 7.06–7.39 (4H, m, aromatics). Anal. calcd for C14H15FN2O4: C, 57.14; H, 5.14; N, 9.52. Found: C, 57.00; H, 5.29; N, 9.33.
Reaction of (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl chloride with N-(4-chlorobenzyl)-O-methyl-hydroxylamine (Kawase, M. et al., J. Chem. Soc. Perkin Trans. 1, 1979, 643–645) as described in the preparation of compound 1-A gave the title amide as white crystals (95% yield): mp 129–130° C. (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.75 (6H, s, CH3), 3.69 (3H, s, OCH3), 4.79 (2H, s, NCH2), 6.39 (1H, s, CH), 7.4 (4H, s, aromatics). Anal. calcd. for C15H16ClNO5: C, 55.31; H, 4.95; N, 4.30. Found: C, 55.32; H, 4.95; N, 4.27.
N-(4-Chlorobenzyl)-2-(2,2-dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-methoxy-acetamide was treated with methanol as described in the preparation of Compound 44-D and gave the title ester as a white syrup (52% yield). 1HNMR 400 MHz (CDCl3) δ (ppm): 3.70 (3H, s, OCH3), 3.90 (3H, s, OCH3), 4.80 (2H, s, NCH2), 6.45 (1H, s, CH), 7.25–7.33 (4H, m, aromatics). Anal. calcd for C13H14ClNO5: C, 52.10; H, 4.71; N, 4.67. Found: C, 51.86; H, 4.68; N, 4.45.
3-[(4-Chlorobenzyl)-methoxy-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 51-B) was treated with paraformaldehyde and methylamine as described in the preparation of Compound 12 to give the title compound as a white solid (69% yield); mp 165° C. dec. 1HNMR 400 MHz (CDCl3) δ (ppm): 3.10 (3H, s, NCH3), 3.72 (3H, s, OCH3), 4.15 (2H, s, NCH2), 4.84 (2H, s, NCH2), 7.25–7.34 (4H, m, aromatics). Anal. calcd for C14H15ClN2O4: C, 54.11; H, 4.87; N, 9.02. Found: C, 53.88; H, 4.71; N, 8.78.
Reaction of 4-fluorophenylacetaldehyde with methoxylamine hydrochloride as described in the preparation of Compound 44-A gave the title oxime ether as a clear oil (43% yield). 1HNMR indicated a 1:1 mixture of E- to Z-isomers. 1HNMR 400 MHz (CDCl3) δ (ppm): 3.51 (2H, d, J=6.7 Hz, CH2), 3.66 (2H, d, J=5.5 Hz, CH2), 3.88 (3H, s, OCH3), 3.96 (3H, s, OCH3), 6.79 (1H, t, J=5.5 Hz, CH), 7.03 (2H, m, aromatics), 7.19 (2H, m, aromatics), 7.45 (1H, t, J=6.7 Hz, CH).
Reduction of 4-fluorophenylacetaldehyde O-methyloxime with sodium cyanoborohydride as described in the preparation of Compound 44-B gave the title hydroxylamine as a clear oil after chromatography on silica gel (62% yield). 1HNMR 400 MHz (C6D6) δ (ppm): 2.64 (2H, d, J=7.1 Hz, CH2), 2.97 (2H, d, J=7.1 Hz, CH2), 3.53 (3H, s, OCH3), 5.24 (broad, NH), 6.9 (4H, m, aromatics).
Reaction of (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl chloride with N-[2-(4-fluorophenyl)-ethyl]-O-methyl-hydroxylamine as described in the preparation of Compound 44-C gave the title amide as white crystals (86% yield): mp 106–107° C. (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.76 (6H, s, CH3), 2.95 (2H, m, CH2), 3.72 (3H, s, OCH3), 3.87 (2H, m, NCH2), 6.38 (1H, broad s, CH), 6.99 (2H, m, aromatics), 7.20 (2H, m, aromatics). Anal. calcd for C16H18FNO5: C, 59.43; H, 5.61; N, 4.33. Found: C, 59.39; H, 5.43; N, 4.13.
2-(2,2-Dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-[2-(4-fluorophenyl)-ethyl]-N-methoxy-acetamide was treated with methanol as described in the preparation Compound 44-D and gave the title ester as a clear oil (66% yield). HRMS (MAB N2) calculated for C14H16FNO5: [M]+: 297.101251. found: 297.101514. 1HNMR 400 MHz (CDCl3) δ (ppm): 2.95 (2H, t, J=7.6 Hz, CH2), 3.73 (3H, s, OCH3), 3.89 (2H, t, J=7.6 Hz, CH2), 3.92 (3H, s, OCH3), 6.44 (1H, s, CH), 7.0 (2H, m, aromatics), 7.19 (2H, m, aromatics).
3-{[2-(4-Fluorophenyl)-ethyl]-methoxy-carbamoyl}-2-hydroxy-acrylic acid methyl ester (Compound 52-D) was treated with paraformaldehyde and methylamine as described in the preparation of Compound 12 to give the title compound as a white solid (38% yield); mp 157° C. dec. 1HNMR 400 MHz (CDCl3) δ (ppm): 2.96 (2H, t, J=7.6 Hz, CH2), 3.14 (3H, s, NCH3), 3.71 (3H, s, OCH3), 3.93 (2H, t, J=7.6 Hz, CH2), 4.14 (2H, s, NCH2), 7.01 (2H, m, aromatics), 7.2 (2H, m, aromatics), 11.55 (1H, broad s, OH). Anal. calcd for C15H17FN2O4: C, 58.44; H, 5.56; N, 9.09. Found: C, 58.52; H, 5.66; N, 8.89.
Reaction of 4-fluorobenzaldehyde with ethoxylamine hydrochloride as described in the preparation of Compound 44-A gave the title oxime ether as a clear oil after chromatography on silica gel (elution toluene-ethyl acetate 95:5) and distillation (58% yield). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.35 (3H, t, J=7.07 Hz, CH3), 4.24 (2H, q, J=7.07 Hz, OCH2), 7.08 (2H, m, aromatics), 7.59 (2H, m, aromatics), 8.07 (1H, s, CH).
Reduction of 4-fluorobenzaldehyde O-ethyloxime with sodium cyanoborohydride as described in the preparation of Compound 44-B gave the title hydroxylamine as a clear oil after chromatography (74% yield). 1HNMR 400 MHz (C6D6) δ (ppm): 1.13 (3H, t, J=7.1 Hz, CH3), 3.70 (2H, q, J=7.1 Hz, OCH2), 3.78 (2H, d, J=5.4 Hz, NCH2), 5.20 (2H, broad t, NH), 6.89 (2H, m, aromatics), 7.09 (2H, m, aromatics). Anal. calcd for C9H12FNO: C, 63.88; H, 7.14; N, 8.27. Found: C, 63.68; H, 7.08; N, 8.46.
Reaction of (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl chloride with O-ethyl-N-4-fluorobenzyl-hydroxylamine as described in the preparation of Compound 44-C gave the title amide as white crystals (92% yield): mp 95–96° C. (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.27 (3H, t, J=7.07 Hz, CH3), 1.77 (6H, s, CH3), 3.90 (2H, q, J=7.07 Hz, OCH2), 4.81 (2H, s, NCH2), 6.41 (1H, s, CH), 7.03 (2H, m, aromatics), 7.37 (2H, m, aromatics). Anal. calcd for C16H18FNO5: C, 59.43; H, 5.61; N, 4.33. Found: C, 59.50; H, 5.60; N, 4.17.
2-(2,2-Dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-ethoxy-N-(4-fluorobenzyl)-acetamide was treated with methanol as described in the preparation of Compound 44-D and gave the title ester as white crystals (84% yield); mp 61–62° C. 1HNMR 400 MHz (CDCl3) δ (ppm): 1.28 (3H, t, J=7.1 Hz, CH3), 3.91 (3H, s, OCH3), 3.92 (2H, q, J=7.1 Hz, OCH2), 4.82 (2H, s, NCH2), 6.47 (1H, s, CH), 7.05 (2H, m, aromatics), 7.32 (2H, m, aromatics), 13.5 (1H, broad s, OH). Anal. calcd for C14H16FNO5: C, 56.56; H, 5.42; N, 4.71. Found: C, 56.67; H, 5.25; N, 4.64.
3-[Ethoxy-(4-fluorobenzyl)-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 53-D) was treated with paraformaldehyde and methylamine as described in the preparation of Compound 12 to give the title compound as a white solid (50% yield); mp 113–114° C. 1HNMR 400 MHz (CDCl3) δ (ppm): 1.30 (3H, t, J=7.1 Hz, CH3), 3.12 (3H, s, NCH3), 3.96 (2H, q, J=7.1 Hz, OCH2), 4.16 (2H, s, NCH2), 4.86 (2H, s, NCH2), 7.06 (2H, m, aromatics), 7.33 (2H, m, aromatics), 11.65 (1H, broad s, OH). Anal. calcd for C15H17FN2O4: C, 58.43; H, 5.55; N, 9.08. Found: C, 58.30; H, 5.55; N, 9.03.
Reaction of 3,4-difluorobenzaldehyde with methoxylamine hydrochloride as described in the preparation of Compound 44-A gave the title oxime ether as a clear oil (100% yield). 1HNMR indicated a 85:15 mixture of E- to Z-isomers. 1HNMR 400 MHz (CDCl3) δ (ppm): (E-isomer) 3.97 (3H, s, OCH3), 7.12–7.26 (2H, m, aromatics), 7.44–7.52 (1H, m, aromatic), 7.97 (1H, s, CH).
Reduction of 3,4-difluorobenzaldehyde O-methyloxime with sodium cyanoborohydride as described in the preparation of Compound 44-B gave the title hydroxylamine as a clear oil (82% yield). 1HNMR 400 MHz (CDCl3) δ (ppm): 3.48 (3H, s, OCH3), 3.98 (2H, s, NCH2), 5.73 (1H, broad s, NH), 7.04–7.23 (3H, m, aromatics). The hydrochloride salt was obtained as a white solid: mp 139–142° C. (dec.). Anal. calcd. for C8H9F2NO2—HCl: C, 45.83; H, 4.80; N, 6.68. Found: C, 45.96; H, 4.93, N, 6.67.
Reaction of (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl chloride with N-3,4-difluorobenzyl-O-methyl-hydroxylamine as described in the preparation of Compound 44-C gave the title amide as a white solid (96% yield): mp 110–111° C. (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.76 (6H, s, CH3), 3.71 (3H, s, OCH3), 4.72 (2H, s, NCH2), 6.38 (1H, s, CH), 7.05–7.22 (3H, m, aromatics). Anal. calcd. for C15H15NO5: C, 55.04; H, 4.62; N, 4.28. Found: C, 54.99; H, 4.55; N, 4.22.
N-(3,4-Difluorobenzyl)-2-(2,2-dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-methoxy-acetamide was treated with methanol as described in the preparation of Compound 44-D and gave the title ester as white crystals (53% yield); mp 76–77° C. 1HNMR 400 MHz (CDCl3) δ (ppm): 3.72 (3H, s, OCH3), 3.90 (3H, s, OCH3), 4.77 (2H, s, NCH2), 6.45 (1H, s, CH), 7.05–7.19 (3H, m, aromatics). Anal. calcd for C13H13F2NO5: C, 51.83; H, 4.35; N, 4.65. Found: C, 51.78; H, 4.28; N, 4.53.
3-[(3,4-Difluorobenzyl)-methoxy-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 54-D) was treated with paraformaldehyde and methylamine as described in the preparation of Compound 12 to and gave the title compound as a white solid (53% yield); mp 146–147° C. 1HNMR 400 MHz (CDCl3) δ (ppm): 3.11 (3H, s, NCH3), 3.74 (3H, s, OCH3), 4.16 (2H, s, NCH2), 4.79 (2H, s, NCH2), 7.06–7.19 (3H, m, aromatics). Anal. calcd for C14H14F2N2O4: C, 53.84; H, 4.51; N, 8.97. Found: C, 53.82; H, 4.41; N, 8.87.
Reaction of 4-methoxybenzaldehyde with methoxylamine hydrochloride as described in the preparation of Compound 44-A gave the title oxime ether as a clear oil (100% yield). 1HNMR indicated a 95:5 mixture of E- to Z-isomers. 1HNMR 400 MHz (CDCl3) δ (ppm): (E-isomer) 3.83 (3H, s, OCH3), 3.94 (3H, s, OCH3), 6.89 (2H, m, aromatics), 7.52 (2H, m, aromatics), 8.05 (1H, s, CH).
Reduction of 4-methoxybenzaldehyde O-methyloxime with sodium cyanoborohydride as described in the preparation of Compound 44-B gave the title hydroxylamine as a clear oil (96% yield). 1HNMR 400 MHz (CDCl3) δ (ppm): 3.49 (3H, s, OCH3), 3.79 (3H, s, OCH3), 3.98 (2H, s, NCH2), 5.62 (1H, broad s, NH), 6.86 (2H, m, aromatics), 7.25 (2H, m, aromatics). The hydrochloride salt was obtained as a white solid: mp 157–158° C. (dec.). Anal. calcd. for C9H13NO2—HCl: C, 53.03; H, 6.92; N, 6.87. Found: C, 53.14; H, 6.76; N, 6.80.
Reaction of (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl chloride with N-(4-methoxybenzyl)-O-methyl-hydroxylamine as described in the preparation of Compound 44-C gave the title amide as white crystals (97% yield): mp 113–114° C. (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.75 (6H, s, CH3), 3.66 (3H, s, OCH3), 3.79 (3H, s, OCH3), 4.77 (2H, s, NCH2), 6.38 (1H, s, CH), 6.85 (2H, m, aromatics), 7.29 (2H, m, aromatics). Anal. calcd. for C16H19NO6: C, 59.80; H, 5.96; N, 4.35. Found: C, 59.87; H, 5.76; N, 4.17.
2-(2,2-Dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-methoxy-N-(4-methoxybenzyl)-acetamide was treated with methanol as described in the preparation of Compound 44-D and gave the title ester as white crystals (56% yield); mp 87–89° C. 1HNMR 400 MHz (CDCl3) δ (ppm): 3.67 (3H, s, OCH3), 3.80 (3H, s, OCH3), 3.89 (3H, s, OCH3), 4.77 (2H, s, NCH2), 6.44 (1H, s, CH), 6.87 (2H, m, aromatics), 7.26 (2H, m, aromatics). Anal. calcd for C14H17NO6: C, 56.94; H, 5.80; N, 4.74. Found: C, 57.03; H, 5.82; N, 4.68.
2-Hydroxy-3-[methoxy-(4-methoxybenzyl)-carbamoyl]-acrylic acid methyl ester (Compound 55-D) was treated with paraformaldehyde and methylamine as described in the preparation of Compound 12 to give the title compound as a white solid (58% yield); mp 135–137° C. 1HNMR 400 MHz (CDCl3) δ (ppm): 3.09 (3H, s, NCH3), 3.71 (3H, s, OCH3), 3.80 (3H, s, OCH3), 4.14 (2H, s, NCH2), 4.82 (2H, s, NCH2), 6.88 (2H, m, aromatics), 7.27 (2H, m, aromatics). Anal. calcd for C15H18N2O5: C, 58.81; H, 5.92; N, 9.14. Found: C, 58.62; H, 5.88; N, 9.12.
Reaction of 2-methylbenzaldehyde with methoxylamine hydrochloride as described in the preparation of Compound 44-A gave the title oxime ether as a clear oil (96% yield). HPLC indicated a 95:5 mixture of E- to Z-isomers. 1HNMR 400 MHz (CDCl3) δ (ppm): (E-isomer) 2.44 (3H, s, CH3), 4.01 (3H, s, OCH3), 7.19–7.28 (3H, m, aromatics), 7.73 (1H, m, aromatic), 8.36 (1H, s, CH).
Reduction of 2-methylbenzaldehyde O-methyloxime with sodium cyanoborohydride as described in the preparation of Compound 44-B gave the title hydroxylamine as a clear oil (83% yield). 1HNMR 400 MHz (CDCl3) δ (ppm): 2.42 (3H, s, CH3), 3.55 (3H, s, OCH3), 4.11 (2H, s, NCH2), 5.64 (1H, s, NH), 7.19–7.32 (4H, m, aromatics). The hydrochloride salt was obtained as a white solid: mp 148–150° C. Anal. calcd. for C9H13NO—HCl: C, 57.60; H, 7.51; N, 7.46. Found: C, 57.59; H, 7.69; N, 7.52.
Reaction of (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl chloride with N-(2-methylbenzyl)-O-methyl-hydroxylamine as described in the preparation of Compound 44-C gave the title amide as white crystals (100% yield): mp 96–97° C. (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.78 (6H, s, CH3), 2.4 (3H, s, CH3), 3.59 (3H, s, OCH3), 4.89 (2H, s, NCH2), 6.44 (1H, s, CH), 7.2–7.28 (4H, m, aromatics). Anal. calcd. for C16H19NO5: C, 62.94; H, 6.27; N, 4.59. Found: C, 62.90; H, 6.21; N, 4.52.
2-(2,2-Dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-methoxy-N-(2-methylbenzyl)-acetamide was treated with methanol as described in the preparation of Compound 44-D and gave the title ester as white crystals (61% yield); mp 80–82° C. 1HNMR 400 MHz (CDCl3) δ (ppm): 2.39 (3H, s, CH3), 3.62 (3H, s, OCH3), 3.92 (3H, s, OCH3), 4.89 (2H, s, NCH2), 6.5 (1H, s, CH), 7.22–7.28 (4H, m, aromatics), 13.5 (1H, broad s, OH). Anal. calcd for C14H17NO5: C, 60.20; H, 6.13; N, 5.01. Found: C, 60.07; H, 5.88; N, 4.84.
2-Hydroxy-3-[methoxy-(2-methylbenzyl)-carbamoyl]-acrylic acid methyl ester (Compound 56-D) was treated with paraformaldehyde and methylamine as described in the preparation of Compound 12 to give the title compound as a white solid (61% yield); mp 153–154° C. 1HNMR 400 MHz (CDCl3) δ (ppm): 2.39 (3H, s, CH3), 3.13 (3H, s, NCH3), 3.67 (3H, s, OCH3), 4.19 (2H, s, NCH2), 4.95 (2H, s, NCH2), 7.2–7.3 (4H, m, aromatics), 11.7 (1H, broad s, OH). Anal. calcd for C15H18N2O4: C, 62.05; H, 6.24; N, 9.65. Found: C, 61.79; H, 6.30; N, 9.58.
Reaction of 3-bromo-4-fluorobenzaldehyde with methoxylamine hydrochloride as described in the preparation of Compound 44-A gave the title oxime ether as a clear oil (95% yield). 1HNMR indicated a 95:5 mixture of E- to Z-isomers. 1HNMR 400 MHz (CDCl3) δ (ppm): (E-isomer) 3.97 (3H, s, OCH3), 7.12 (1H, m, aromatics), 7.48 (1H, m, aromatic), 7.82 (1H, m, aromatic), 7.97 (1H, s, CH).
Reduction of 3-bromo-4-fluorobenzaldehyde O-methyloxime with sodium cyanoborohydride as described in the preparation of Compound 44-B gave the title hydroxylamine as a clear oil (83% yield). 1HNMR 400 MHz (CDCl3) δ (ppm): 3.48 (3H, s, OCH3), 3.99 (2H, s, NCH2), 7.08 (1H, m, aromatic), 7.27 (1H, m, aromatic), 7.57 (1H, m, aromatic). The hydrochloride salt was obtained as a white solid: mp 150–151° C. Anal. calcd. for C8H9BrFNO—HCl: C, 35.52; H, 3.73; N, 5.18. Found: C, 35.54; H, 3.61; N, 5.12.
Reaction of (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl chloride with N-3-bromo-4-fluorobenzyl-O-methyl-hydroxylamine as described in the preparation of Compound 44-C gave the title amide as a white solid (100% yield): mp 117–119° C. (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.75 (6H, s, CH3), 3.71 (3H, s, OCH3), 4.76 (2H, s, NCH2), 6.38 (1H, s, CH), 7.07 (1H, m, aromatic), 7.28 (1H, m, aromatic), 7.56 (1H, m, aromatic). Anal. calcd. for C15H15BrFNO5: C, 46.41; H, 3.89; N, 3.61. Found: C, 46.43; H, 4.01; N, 3.53.
N-(3-Bromo-4-fluorobenzyl)-2-(2,2-dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-methoxy-acetamide was treated with methanol as described in the preparation of Compound 44-D and gave the title ester as white crystals (44% yield); mp 107–108° C. 1HNMR 400 MHz (CDCl3) δ (ppm): 3.72 (3H, s, OCH3), 3.90 (3H, s, OCH3), 4.77 (2H, s, NCH2), 6.45 (1H, s, CH), 7.09 (1H, m, aromatic), 7.25 (1H, m, aromatic), 7.53 (1H, m, aromatic). Anal. calcd for C13H13BrFNO5: C, 43.11; H, 3.62; N, 3.86. Found: C, 43.10; H, 3.54; N, 3.87.
3-[(3-Bromo-4-fluorobenzyl)-methoxy-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 57-D) was treated with paraformaldehyde and methylamine as described in the preparation of Compound 12 to give the title compound as a white solid (35% yield); mp 154–157° C. dec. 1HNMR 400 MHz (CDCl3) δ (ppm): 3.11 (3H, s, NCH3), 3.75 (3H, s, OCH3), 4.15 (2H, s, NCH2), 4.81 (2H, s, NCH2), 7.1 (1H, m, aromatic), 7.27 (1H, m, aromatic), 7.54 (1H, m, aromatic). Anal. calcd for C14H14BrFN2O4: C, 45.06; H, 3.78; N, 7.50. Found: C, 44.80; H, 3.81; N, 7.33.
Reaction of 4-fluorobenzaldehyde with O-isobutyl-hydroxylamine hydrochloride as described in the preparation of Compound 44-A gave the title oxime ether as a clear oil after chromatography on silica gel (elution toluene-ethyl acetate 95:5), (77% yield). 1HNMR 400 MHz (CDCl3) δ (ppm): 0.98 (6H, d, J=6.5 Hz, CH3), 2.07 (1H, m, CH), 3.95 (2H, d, J=7.18 Hz, OCH2), 7.08 (2H, m, aromatics), 7.59 (2H, m, aromatics), 8.08 (1H, s, CH). Anal. calcd for C11H14FNO: C, 67.67; H, 7.22; N, 7.17. Found: C, 67.71; H, 7.32; N, 7.38.
Reduction of 4-fluorobenzaldehyde O-isobutyloxime with sodium cyanoborohydride as described in the preparation of Compound 44-B gave the title hydroxylamine as a clear oil after chromatography (65% yield). 1HNMR 400 MHz (C6D6) δ (ppm): 0.87 (6H, d, J=6.75 Hz, CH3), 1.88 (1H, m, CH), 3.46 (2H, d, J=6.41 Hz, OCH2), 4.05 (2H, s, NCH2), 7.04 (2H, m, aromatics), 7.37 (2H, m, aromatics). Anal. calcd for C11H16FNO: C, 66.98; H, 8.17; N, 7.10. Found: C, 66.88; H, 7.97; N, 7.32.
Reaction of (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl chloride with N-(4-fluorobenzyl)-O-isobutyl-hydroxylamine as described in the preparation of Compound 44-C gave the title amide as white crystals (91% yield): mp 105–106° C. (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ (ppm): 0.98 (3H, d, J=6.45 Hz, CH3), 1.77 (6H, s, CH3), 1.95 (1H, m, CH), 3.64 (2H, d, J=6.63 Hz, OCH2), 4.80 (2H, s, NCH2), 6.41 (1H, s, CH), 7.03 (2H, m, aromatics), 7.36 (2H, m, aromatics). Anal. calcd for C18H22FNO5: C, 61.53; H, 6.31; H, 3.98. Found: C, 61.47; H, 6.39; N, 3.97.
2-(2,2-Dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-(4-fluorobenzyl)-N-isobutoxy-acetamide was treated with methanol as described in the preparation of Compound 44-D and gave the title ester as white crystals (48% yield); mp 55–56° C. 1HNMR 400 MHz (CDCl3) δ (ppm): mixture of rotamers: 0.99 (6H, d, J=7.1 Hz, CH3), 1.96 (1H, m, CH), 3.65 (2H, d, J=6.0 Hz, OCH2), 3.91 (3H, s, OCH3), 4.81 (2H, s, NCH2), 6.49 (1H, s, CH), 7.05 (2H, m, aromatics), 7.33 (2H, m, aromatics), 13.4 (1H, broad s, OH). HRMS (ES+) calculated for C16H21FNO5, [M+H]+: 326.140376. found: 326.139560.
3-[(4-Fluorobenzyl)-isobutoxy-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 58-D) was treated with paraformaldehyde and methylamine as described in the preparation of Compound 12 to give the title compound as a white solid (40% yield); mp 96–97° C. 1HNMR 400 MHz (CDCl3) δ (ppm): 0.98 (6H, d, J=6.5 Hz, CH3), 1.96 (1H, m CH), 3.12 (3H, s, NCH3), 3.69 (2H, d, J=6.4 Hz, OCH2), 4.1 (2H, s, NCH2), 4.85 (2H, s, NCH2), 7.06 (2H, m, aromatics), 7.33 (2H, m, aromatics), 11.65 (1H, broad s, OH). HRMS (ES+) calculated for C17H22FN2O4, [M+H]+: 337.156361. found: 337.156153.
2-(2,2-Dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-methoxy-N-(4-methylbenzyl)-acetamide, prepared using the methods described in the previous examples, was treated with methanol as described in the preparation of Compound 44-D to give the title ester as white crystals (69% yield); mp 83–84° C. 1HNMR 400 MHz (CDCl3) δ (ppm): 2.59 (3H, s, CH3), 3.94 (3H, s, OCH3), 4.15 (3H, s, OCH3), 5.05 (2H, s, NCH2), 6.72 (1H, s, CH), 7.41 (2H, d, J=8.1 Hz, aromatics), 7.47 (2H, d, J=8.1 Hz, aromatics), 13.75 (1H, broad s, OH). Anal. calcd for C14H17NO5: C, 60.20; H, 6.13; N, 5.01. Found: C, 60.24; H, 6.09; N, 4.85.
2-Hydroxy-3-[methoxy-(4-methylbenzyl)-carbamoyl]-acrylic acid methyl ester (Compound 59-A) was treated with paraformaldehyde and methylamine as described in the preparation of Compound 12 to give the title compound as a white solid (41% yield); mp 150–152° C. 1HNMR 400 MHz (CDCl3) δ (ppm): 2.4 (3H, s, CH3), 3.16 (3H, s, NCH3), 3.78 (3H, s, OCH3), 4.21 (2H, s, NCH2), 4.91 (2H, s, NCH2), 7.22 (2H, d, J=8 Hz, aromatics), 7.28 (2H, d, J=8 Hz, aromatics), 11.7 (1H, broad s, OH). Anal. calcd for C15H18N2O4: C, 62.05; H, 6.24; N, 9.65. Found: C, 61.91; H, 6.30; N, 9.56.
Reaction of 2,4-difluorobenzaldehyde with methoxylamine hydrochloride as described in the preparation of Compound 44-A gave the title oxime ether as a clear oil (80% yield). 1HNMR indicated a 95:5 mixture of E- and Z-isomers. 1HNMR 400 MHz (CDCl3) δ (ppm): (E-isomer) 3.98 (3H, s, OCH3), 6.79–6.91 (2H, m, aromatics), 7.79–7.85 (1H, m, aromatic), 8.24 (1H, s, CH).
Reduction of 2,4-difluorobenzaldehyde O-methyloxime with sodium cyanoborohydride as described in the preparation of Compound 44-B gave the title hydroxylamine as a clear oil (72% yield). 1HNMR 400 MHz (CDCl3) δ (ppm): 3.51 (3H, s, OCH3), 4.07 (2H, s, NCH2), 6.78–6.88 (2H, m, aromatics), 7.32–7.38 (1H, m, aromatic). The hydrochloride salt was obtained as a white solid: mp 154–158° C. (dec.). Anal. calcd. for C8H9NO2—HCl: C, 45.83; H, 4.80; N, 6.68. Found: C, 45.81; H, 4.84; N, 6.59.
Reaction of (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl chloride with N-2,4-difluorobenzyl-O-methyl-hydroxylamine as described in the preparation of Compound 44-C gave the title amide as a white solid (97% yield): mp 120–125° C. (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.75 (6H, s, CH3), 3.73 (3H, s, OCH3), 4.86 (2H, s, NCH2), 6.38 (1H, s, CH), 6.78–6.87 (2H, m, aromatics), 7.37–7.43 (1H, m, aromatic). Anal. calcd. for C15H15F2NO5: C, 55.04; H, 4.62; N, 4.28. Found: C, 55.03; H, 4.43; N, 4.17.
N-(2,4-Difluorobenzyl)-2-(2,2-dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-methoxy-acetamide was treated with methanol as described in the preparation of Compound 44-D and gave the title ester as white crystals (55% yield); mp 104–105° C. 1HNMR 400 MHz (CDCl3) δ (ppm): 3.73 (3H, s, OCH3), 3.89 (3H, s, OCH3), 4.87 (2H, s, NCH2), 6.45 (1H, s, CH), 6.8–6.9 (2H, m, aromatics), 7.31–7.37 (1H, m, aromatic). Anal. calcd for C13H13F2NO5: C, 51.83; H, 4.35; N, 4.65. Found: C, 51.68; H, 4.27; N, 4.53.
3-[(2,4-Difluorobenzyl)-methoxy-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 60-D) was treated with paraformaldehyde and methylamine as described in the preparation of Compound 12 to give the title compound as a white solid (55% yield); mp 141–149° C., dec. 1HNMR 400 MHz (CDCl3) δ (ppm): 3.1 (3H, s, NCH3), 3.77 (3H, s, OCH3), 4.14 (2H, s, NCH2), 4.92 (2H, s, NCH2), 6.8–6.9 (2H, m, aromatics), 7.35–7.41 (1H, m, aromatic).
Reaction of 3-cyano-4-fluorobenzaldehyde with methoxylamine hydrochloride as described in the preparation of Compound 44-A gave the title oxime ether as a clear oil after chromatography on silica gel (elution hexane-ethyl acetate 8:2) (94% yield). 1HNMR indicated a 93:7 mixture of E- to Z-isomers. 1HNMR 400 MHz (CDCl3) δ (ppm): (E-isomer) 4.02 (3H, s, OCH3), 7.26 (1H, m, aromatic), 7.85 (2H, m, aromatics), 8.03 (1H, s, CH).
Reduction of 3-cyano-4-fluorobenzaldehyde O-methyloxime with sodium cyanoborohydride as described in the preparation of Compound 44-B gave the title hydroxylamine as a clear oil after chromatography on silica gel (elution hexane-ethyl acetate 8:2) (73% yield). 1HNMR 400 MHz (CDCl3) δ (ppm): 3.46 (3H, s, OCH3), 4.02 (2H, s, NCH2), 7.18 (1H, t, aromatic), 7.58–7.66 (2H, m, aromatics). The hydrochloride salt was obtained as a white solid: mp 152–158° C. Anal. calcd for C9H9FN2O—HCl: C, 49.89; H, 4.65; N, 12.93. Found: C, 50.04; H, 4.64; N, 12.84.
Reaction of (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl chloride with N-(3-cyano-4-fluorobenzyl)-O-methyl-hydroxylamine as described in the preparation of Compound 44-C gave the title amide as white crystals (97% yield): mp 119–120° C. (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.75 (6H, s, CH3), 3.75 (3H, s, OCH3), 4.78 (2H, s, NCH2), 6.36 (1H, s, CH), 7.17 (1H, t, aromatic), 7.58–7.64 (2H, m, aromatics). Anal. calcd for C16H15F2NO5: C, 57.48; H, 4.52; N, 8.38. Found: C, 57.39; H, 4.61; N, 8.32.
N-(3-Cyano-4-fluorobenzyl)-2-(2,2-dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-methoxy-acetamide was treated with methanol as described in the preparation of Compound 44-D and gave the title ester as white crystals (47% yield); mp 125–126° C. 1HNMR 400 MHz (CDCl3) δ (ppm): 3.75 (3H, s, OCH3), 3.90 (3H, s, OCH3), 4.81 (2H, s, NCH2), 6.44 (1H, s, CH), 7.18 (1H, m, aromatic), 7.56–7.61 (2H, m, aromatics). Anal. calcd for C14H13FN2O5: C, 54.54; H, 4.25; N, 9.08. Found: C, 54.76; H, 4.29; N, 9.04.
3-[(3-Cyano-4-fluorobenzyl)-methoxy-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 61-D) was treated with paraformaldehyde and methylamine as described in the preparation of Compound 12 to give the title compound as a white solid (69% yield); mp 175° C. dec. 1HNMR 400 MHz (CDCl3) δ (ppm): 3.11 (3H, s, NCH3), 3.78 (3H, s, OCH3), 4.16 (2H, s, NCH2), 4.85 (2H, s, NCH2), 7.19–7.24 (1H, m, aromatic), 7.59–7.62 (2H, m, aromatics).
Reaction of 4-cyanobenzaldehyde with methoxylamine hydrochloride as described in the preparation of Compound 44-A gave the title oxime ether as a white solid (96% yield), (Gordon, M. S. et al., J. Org. Chem., 49, 1984, 97–100). 1HNMR indicated a 95:5 mixture of E- to Z-isomers. 1HNMR 400 MHz (CDCl3) δ (ppm): (E-isomer) 4.02 (3H, s, OCH3), 7.07 (4H, m, aromatics), 8.06 (1H, s, CH).
Reduction of 4-cyanobenzaldehyde O-methyloxime with sodium cyanoborohydride as described in the preparation of Compound 44-B gave the title hydroxylamine as a clear oil (75% yield). 1HNMR 400 MHz (CDCl3) δ (ppm): 3.48 (3H, s, OCH3), 4.09 (2H, s, NCH2), 7.48 (2H, m, aromatics), 7.63 (2H, m, aromatics). The hydrochloride salt was obtained as a white solid: mp 168° C. (dec.). Anal. calcd. for C9H10N2O—HCl: C, 54.41; H, 5.58; N, 14.10. Found: C, 54.44; H, 5.62; N, 13.94.
Reaction of (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl chloride with N-(4-cyanobenzyl)-O-methyl-hydroxylamine as described in the preparation of Compound 44-C gave the title amide as white crystals (99% yield): mp 148–149° C. (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.75 (6H, s, CH3), 3.72 (3H, s, OCH3), 4.86 (2H, s, NCH2), 6.39 (1H, s, CH), 7.46 (2H, m, aromatics), 7.63 (2H, m, aromatics). Anal. calcd. for C16H16N2O5: C, 60.75; H, 5.10; N, 8.86. Found: C, 60.60; H, 4.91; N, 8.78.
N-(4-Cyanobenzyl)-2-(2,2-dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-methoxy-acetamide was treated with methanol as described in the preparation of Compound 44-D gave the title ester as white crystals (52% yield); mp 110° C. 1HNMR 400 MHz (CDCl3) δ (ppm): 3.73 (3H, s, OCH3), 3.90 (3H, s, OCH3), 4.88 (2H, s, NCH2), 6.47 (1H, s, CH), 7.43 (2H, d, J=8.6 Hz, aromatics), 7.64 (2H, d, J=8.6 Hz, aromatics). Anal. calcd for C14H14N2O5: C, 57.93; H, 4.86; N, 9.65. Found: C, 57.87; H, 4.80; N, 9.67.
3-[(4-Cyanobenzyl)-methoxy-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 62-D) was treated with paraformaldehyde and methylamine as described in the preparation of Compound 12 to give the title compound as a white solid (46% yield); mp 161° C. dec. 1HNMR 400 MHz (CDCl3) δ (ppm): 3.11 (3H, s, NCH3), 3.75 (3H, s, OCH3), 4.17 (2H, s, NCH2), 4.93 (2H, s, NCH2), 7.44 (2H, d, J=8.2 Hz, aromatics), 7.65 (2H, d, J=8.2 Hz, aromatics). Anal. calcd for C15H15N3O4: C, 59.79; H, 5.01; N, 13.94. Found: C, 59.51; H, 4.90; N, 13.69.
Reaction of 4-acetamidobenzaldehyde with methoxylamine hydrochloride as described in the preparation of Compound 44-A gave the title oxime ether as a white solid (98% yield), (Sakamoto, T. et al., J. Org. Chem., 57, 1992, 3245–3248). 1HNMR indicated a 95:5 mixture of E- to Z-isomers. 1HNMR 400 MHz (CDCl3) δ (ppm): (E-isomer) 2.19 (3H, s, CH3), 3.96 (3H, s, OCH3), 7.22 (1H, broad s, NH), 7.53 (4H, m, aromatics), 8.01 (1H, s, CH).
Reduction of 4-acetamidobenzaldehyde O-methyloxime with sodium cyanoborohydride as described in the preparation of Compound 44-B gave the title hydroxylamine as a waxy solid (100% yield). 1HNMR 400 MHz (CDCl3) δ (ppm): 2.16 (3H, s, CH3), 3.49 (3H, s, OCH3), 4.00 (2H, s, NCH2), 7.26 (1H, broad s, NH), 7.29 (2H, m, aromatics), 7.46 (2H, m, aromatics). The hydrochloride salt was obtained as a white solid: mp 186–188° C. (dec.). Anal. calcd. for C10H14N2O2—HCl—H2O: C, 50.87; H, 6.66; N, 11.87. Found: C, 50.77; H, 6.44; N, 12.16.
Reaction of (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl chloride with N-(4-acetamidobenzyl)-O-methyl-hydroxylamine as described in the preparation of Compound 44-C gave the title amide as white crystals (92% yield): mp 212–215° C. (dec.) (dichloromethane-hexane). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.73 (6H, s, CH3), 2.16 (3H, s, CH3), 3.67 (3H, s, OCH3), 4.78 (2H, s, NCH2), 6.39 (1H, s, CH), 7.32 (3H, m, aromatics and NH), 7.45 (2H, m, aromatics). Anal. calcd. for C17H20N2O6: C, 57.87; H, 5.86; N, 7.94. Found: C, 57.76; H, 5.68; N, 8.51.
N-(4-Acetylaminobenzyl)-2-(2,2-dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-methoxy-acetamide was treated with methanol as described in the preparation of Compound 44-D and gave the title ester as white crystals (35% yield); mp 125° C. 1HNMR 400 MHz (CDCl3) δ (ppm): 2.18 (3H, s, COCH3), 3.68 (3H, s, OCH3), 3.89 (3H, s, OCH3), 4.79 (2H, s, NCH2), 6.46 (1H, s, CH), 7.16 (1H broad s, NH), 7.29 (2H, d, J=8.6 Hz, aromatics), 7.48 (2H, d, J=8.6 Hz, aromatics). Anal. Calcd for C15H18N2O6: C, 55.89; H, 5.62; N, 8.69. Found: C, 55.95; H, 5.70; N, 8.59.
3-[(4-Acetylaminobenzyl)-methoxy-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 63-D) was treated with paraformaldehyde and methylamine as described in the preparation Compound 12 to give the title compound as a white solid (43% yield); mp 110° C. 1HNMR 400 MHz (CDCl3) δ (ppm): 2.18 (3H, s, COCH3), 3.1 (3H, s, NCH3), 3.71 (3H, s, OCH3), 4.15 (2H, s, NCH2), 4.83 (2H, s, NCH2), 7.29 (2H, d, J=8.6 Hz, aromatics), 7.34 (1H, broad s, NH), 7.50 (2H, d, J=8.6 Hz, aromatics). HRMS (ES+) calculated for C16H20N3O5: [M+H]+: 334.140296. found: 334.139137.
2-(2,2-Dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-(4-fluoro-3-methylbenzyl)-N-methoxy-acetamide, prepared using the methods described in the previous examples, was treated with methanol as described in the preparation of Compound 44-C and gave the title ester as white crystals (32% yield); mp 60–62° C. 1HNMR 400 MHz (CDCl3) δ (ppm): 2.27 (3H, broad s, CH3), 3.70 (3H, s, OCH3), 3.90 (3H, s, OCH3), 4.76 (2H, s, NCH2), 6.46 (1H, s, CH), 6.96 (1H, m, aromatic), 7.12 (2H, m, aromatics). Anal. calcd for C14H16FNO5: C, 56.56; H, 5.42; N, 4.71. Found: C, 56.36; H, 5.44; N, 4.54.
3-[(4-Fluoro-3-methylbenzyl)-methoxy-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 64-A) was treated with paraformaldehyde and methylamine as described in the preparation of Compound 12 to give the title compound as white crystals (76% yield); mp 160–164° C. 1HNMR 400 MHz (CDCl3) δ (ppm): 2.27 (3H, s, CH3), 3.10 (3H, s, NCH3), 3.73 (3H, s, OCH3), 4.15 (2H, s, NCH2), 4.81 (2H, s, NCH2), 6.97 (1H, m, aromatic), 7.12 (2H, m, aromatics). Anal. calcd for C15H17FN2O4: C, 58.44; H, 5.56; N, 9.09. Found: C, 58.41; H, 5.61; N, 8.90.
2-(2,2-Dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-(3-fluoro-4-methylbenzyl)-N-methoxy-acetamide, prepared using the methods described in the previous examples, was treated with methanol as described in the preparation of Compound 44-C and gave the title ester as white crystals (49% yield); mp 88° C. 1HNMR 400 MHz (CDCl3) δ (ppm): 2.25 (3H, d, J=1.5 Hz, CH3), 3.70 (3H, s, OCH3), 3.90 (3H, s, OCH3), 4.78 (2H, s, NCH2), 6.47 (1H, s, CH), 6.96 (2H, m, aromatics), 7.26 (1H, m, aromatic). HRMS (MAB N2) calculated for C14H16FNO5, [M]+: 297.101251. found: 297.101261.
3-[(3-Fluoro-4-methylbenzyl)-methoxy-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 65-A) was treated with paraformaldehyde and methylamine as described in the preparation of Compound 12 to give the title compound as white crystals (54% yield); mp 145° C. 1HNMR 400 MHz (CDCl3) δ (ppm): 2.26 (3H, broad s, CH3), 3.11 (3H, s, NCH3), 3.73 (3H, s, OCH3), 4.16 (2H, s, NCH2), 4.83 (2H, s, NCH2), 6.99 (2H, m, aromatics), 7.16 (1H, m, aromatic). Anal. calcd for C15H17FN2O4: C, 58.44; H, 5.56; N, 9.09. Found: C, 58.16; H, 5.44; N, 8.88.
2-(2,2-Dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-(2-isopropoxybenzyl)-N-methoxy-acetamide, prepared using the methods described in the previous examples, was treated with methanol as described in the preparation of Compound 44-D and gave the title ester as a white syrup (62% yield). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.33 (6H, d, J=6 Hz, CH3), 3.68 (3H, s, OCH3), 3.89 (3H, s, OCH3), 4.59 (1H, m, CH), 4.9 (2H, s, NCH2), 6.50 (1H, s, CH), 6.88 (2H, m, aromatics), 7.24 (2H, m, aromatics). HRMS (MAB N2) calculated for C16H21NO6, [M]+: 323.136888. found: 323.136700.
2-Hydroxy-3-[(2-isopropoxybenzyl)-methoxy-carbamoyl]-acrylic acid methyl ester (Compound 66-A) was treated with paraformaldehyde and methylamine as described in the preparation of Compound 12 to give the title compound as a tan solid (22% yield). 1HNMR 400 MHz (DMSO-d6) δ (ppm): 1.26 (6H, d, J=6.2 Hz, CH3), 2.91 (3H, s, NCH3), 3.57 (3H, s, OCH3), 4.03 (2H, s, NCH2), 4.60 (1H, m, CH), 4.71 (2H, s, NCH2), 6.66 (1H, m, aromatic), 6.95 (1H, m, aromatic), 7.15 (2H, m, aromatics). HRMS (ES+) calculated for C7H23N2O5, [M+H]+: 335.160697. found: 335.161171.
Compound 67-A: 4-Carbomethoxybenzaldehyde O-methyloxime
Reaction of methyl 4-formylbenzoate with methoxylamine hydrochloride as described in the preparation of Compound 44-A gave the title oxime ether (96% yield) as a white solid (mixture of E- and Z-isomers). The E-isomer was obtained as white crystals from hexane; mp 66–67° C. (Lit. mp 65–66° C., Cooks, Org. Mass Spectrum., 5, 1971, 687). 1HNMR 400 MHz (DMSO-d6) δ (ppm): (E-isomer) 3.86 (3H, s, OCH3), 3.93 (3H, s, OCH3), 7.75 (2H, d, aromatics), 7.98 (2H, d, aromatics), 8.32 (1H, s, CH).
Reduction of 4-carbomethoxybenzaldehyde O-methyloxime with sodium cyanoborohydride as described in the preparation of Compound 44-B gave the title hydroxylamine as an oil (53% yield). The hydrochloride salt was obtained as a white solid: mp 166–169° C. 1HNMR 400 MHz (DMSO-d6) δ (ppm): 3.75 (3H, s, OCH3), 3.86 (3H, s, OCH3), 4.39 (2H, s, NCH2), 7.65 (2H, d, aromatics), 7.97 (2H, d, aromatics). Anal. calcd for C10H13NO3—HCl: C, 51.84; H, 6.09; N, 6.04. Found: C, 51.74; H, 6.01; N, 5.50.
Reaction of (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl chloride with N-4-carbomethoxybenzyl-O-methyl-hydroxylamine as described in the preparation of Compound 44-C gave the title amide as a white solid (83% yield): mp 120° C. (dichloromethane-hexane). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.75 (6H, s, CH3), 3.67 (3H, s, OCH3), 3.91 (3H, s, OCH3), 4.88 (2H, s, NCH2), 6.40 (1H, s, CH), 7.42 (2H, d, aromatics), 8.0 (2H, d, aromatics). Anal. calcd for C17H19NO7: C, 58.45; H, 5.48; N, 4.01. Found: C, 58.54; H, 5.55; N, 3.61.
4-({[2-(2,2-Dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-acetyl]-methoxyamino}-methyl)-benzoic acid methyl ester was treated with methanol as described in the preparation of Compound 44-D and gave the title ester as a white solid. 1HNMR 400 MHz (CDCl3) δ (ppm): 3.70 (3H, s, OCH3), 3.90 (3H, s, OCH3), 3.91 (3H, s, OCH3), 4.89 (2H, s, NCH2), 6.5 (1H, s, CH), 7.39 (2H, d, J=8.1 Hz, aromatics), 8.02 (2H, d, J=8.1 Hz, aromatics). HRMS (ES+) calculated for C15H18NO7, [M+H]+: 324.108327. found: 324.109066.
4-{[(3-Hydroxy-3-[methoxycarbonyl-acryloyl)-methoxyamino]-methyl}-benzoic acid methyl ester (Compound 67-D) was treated with paraformaldehyde and methylamine as described in the preparation of Compound 12 to give the title compound as a tan solid. 1HNMR 400 MHz (CDCl3) δ (ppm): 3.11 (3H, s, NCH3), 3.73 (3H, s, OCH3), 3.92 (3H, s, OCH3), 4.17 (2H, s, NCH2), 4.93 (2H, s, NCH2), 7.4 (2H, d, J=8.1 Hz, aromatics), 8.02 (2H, d, J=8.1 Hz, aromatics), 11.54 (1H, broad s, OH). HRMS (ES+) calculated for C16H19N2O6, [M+H]+: 335.1249. found: 335.1243.
4-({[2-(2,2-dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-acetyl]-methoxyamino}-methyl)-benzoic acid tert-butyl ester, prepared using the methods described in the previous examples, was treated in methanol with paraformaldehyde and methylamine as described in the preparation of Compound 12 to give the title compound as a white solid (40% yield); mp 157–160° C. 1HNMR 400 MHz (CDCl3) δ (ppm): 1.58 (9H, s, t-Bu), 3.11 (3H, s, NCH3), 3.72 (3H, s, OCH3), 4.16 (2H, s, NCH2), 4.92 (2H, s, NCH2), 7.37 (2H, d, J=8 Hz, aromatics), 7.97 (2H, d, J=8 Hz, aromatics), 11.55 (1H, broad s, OH).
A solution of 4-{[(4-hydroxy-1-methyl-5-oxo-2,5-dihydro-1H-pyrrole-3-carbonyl)-methoxyamino]-methyl}-benzoic acid tert-butyl ester (0.062 g, 0.16 mmol) in dichloromethane (3 ml) was treated with trifluoroacetic acid (0.6 ml) and stirred at 25° C. for 2 h. The solvent was then evaporated in vacuo and the residue was triturated with acetonitrile to give 0.041 g (80% yield) of the title material as a white solid; mp 196–197° C. 1HNMR 400 MHz (DMSO-d6) δ (ppm): 2.97 (3H, s, NCH3), 3.73 (3H, s, OCH3), 4.19 (2H, s, NCH2), 4.96 (2H, s, NCH2), 7.42 (2H, d, J=8.3 Hz, aromatics), 7.91 (2H, d, J=8.3 Hz, aromatics), 11.4 (1H, broad s, OH), 12.9 (1H, broad s, OH). Anal. calcd for C15H16N2O6.H2O: C, 53.25; H, 5.36; N, 8.28. Found: C, 53.59; H, 4.79; N, 8.19.
2-(2,2-Dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-[1-(4-fluorophenyl)-ethyl]-N-methoxy-acetamide, prepared using the methods described in the previous examples, was treated in methanol with paraformaldehyde and methylamine as described in the preparation Compound 12 to give the title compound as a white solid (47% yield); mp 136–138° C. 1HNMR 400 MHz (CDCl3) δ (ppm): 1.68 (3H, d, J=7.1 Hz, CH3), 3.1 (3H, s, NCH3), 3.49 (3H, s, OCH3), 4.13 (2H, AB system, JAB=18.2 Hz, Δv=45.7 Hz, NCH2), 5.75 (1H, q, J=7.1 Hz, CH), 7.06 (2H, m, aromatics), 7.42 (2H, m, aromatics). Anal. calcd for C15H17FN2O4: C, 58.43; H, 5.55; N, 9.08. Found: C, 58.40; H, 5.38; N, 9.01.
Condensation of 4-fluorobenzaldehyde with hydroxylamine hydrochloride using the same procedure as Compound 44-A followed by reaction with tert-butyl bromoacetate gave the title oxime ether as a clear oil (84% yield). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.52 (9H, s, t-Bu), 4.61 (2H, s, OCH2), 7.08 (2H, m, aromatics), 7.59 (2H, m, aromatics), 8.19 (1H, s, CH).
Reduction of (4-fluorobenzylideneaminooxy)-acetic acid tert-butyl ester as described in the preparation of Compound 44-B gave the title hydroxylamine as a clear oil (65% yield). 1HNMR 400 MHz (C6D6) δ (ppm): 1.43 (9H, s, t-Bu), 3.84 (2H, d, J=5.6 Hz, NCH2), 4.17 (2H, s, OCH2), 6.39 (1H, broad t, NH), 6.86 (2H, m, aromatics), 7.05 (2H, m, aromatics).
Reaction of (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl chloride with [N-(4-fluorobenzyl)aminooxy]-acetic acid tert-butyl ester as described in the preparation of compound 1-A gave the title amide as white crystals (85% yield): mp 119–120° C. (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.48 (9H, s, t-Bu), 1.74 (6H, s, CH3), 4.30 (2H, s, CH2), 4.88 (2H, s, CH2), 6.48 (1H, s, CH), 7.0 (2H, m, aromatics), 7.38 (2H, m, aromatics). Anal. calcd for C20H24FNO7: C, 58.67; H, 5.91; N, 3.42. Found: C, 58.83; H, 5.92; N, 3.31.
[[2-(2,2-Dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-acetyl]-(4-fluorobenzyl)-aminooxy]-acetic acid tert-butyl ester was treated with methanol as described in the preparation of Compound 44-D and gave the title ester as a clear oil (69% yield). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.50 (9H, s, t-Bu), 3.92 (3H, s, OCH3), 4.35 (2H, s, CH2), 4.94 (2H, s, CH2), 6.55 (1H, s, CH), 7.05 (2H, m, aromatics), 7.39 (2H, m, aromatics), 13.35 (1H, broad s, OH). HRMS (ES+) calculated for C18H23FNO7, [M+H]+: 384.145856. found: 384.146214.
3-[tert-Butoxycarbonylmethoxy-(4-fluorobenzyl)-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 70-D) was treated with paraformaldehyde and methylamine as described in the preparation of Compound 12 to give the title compound as a white solid (30% yield); mp 128–130° C. 1HNMR 400 MHz (CDCl3) δ (ppm): 1.48 (9H, s, t-Bu), 3.1 (3H, s, NCH3), 4.22 (2H, s, CH2), 4.35 (2H, s, CH2), 4.93 (2H, s, NCH2), 7.06 (2H, m, aromatics), 7.38 (2H, m, aromatics), 11.55 (1H, broad s, OH). HRMS (ES+) calculated for C19H24FN2O6, [M+H]+: 395.161840. found: 395.161599.
A solution of [(4-fluorobenzyl)-(4-hydroxy-1-methyl-5-oxo-2,5-dihydro-1H-pyrrole-3-carbonyl)-aminooxy]-acetic acid tert-butyl ester (0.041 g, 0.104 mmol) in dichloromethane (4 ml) was treated with trifluoroacetic acid (1 ml) and the resulting mixture was stirred at 22° C. for 2 h. The solvent was evaporated in vacuo and the residue was recrystallized from a mixture of ethyl acetate and hexane to give 0.027 g (80% yield) of the tittle material as white crystals; mp 147–150° C. 1HNMR 400 MHz (DMSO-d6) δ (ppm): 2.95 (3H, s, NCH3), 4.14 (2H, s, CH2), 4.62 (2H, s, CH2), 4.92 (2H, s, CH2), 7.17 (2H, m, aromatics), 7.36 (2H, m, aromatics), 11.4 (1H, broad s, OH), 13.1 (1H, broad s, OH). HRMS (ES+) calculated for C15H16FN2O6, [M+H]+: 339.099240. found: 339.100624.
A solution of [[2-(2,2-dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-acetyl]-(4-fluorobenzyl)-aminooxy]-acetic acid (0.681 g, 1.93 mmol) in dichloromethane (20 ml) was treated at 22° C. with oxalyl chloride (0.34 ml, 3.9 mmol) and a trace of N,N-dimethylformamide and the resulting mixture was stirred for 1 h. The solvent and excess reagent were then evaporated in vacuo. The residual material was dissolved in dry dichloromethane (10 ml) and added dropwise to a cold (0° C.) solution of dimethylamine (0.18 g, 4.0 mmol) and pyridine (0.25 ml, 3.2 mmol) in dichloromethane. After 2 h, the reaction mixture was diluted with ethyl acetate, washed with saturated sodium bicarbonate and brine and dried over anhydrous magnesium sulfate. Evaporation of the solvent in vacuo and crystallisation of the residue from a mixture of ethyl acetate and hexane gave 0.370 g (50% yield) of the title material as a white solid. 1HNMR 400 MHz (CDCl3) δ (ppm): 1.77 (6H, s, CH3), 2.91 (3H, s, CH3), 2.97 (3H, s, CH3), 4.53 (2H, s, CH2), 4.93 (2H, s, CH2), 6.43 (1H, s, CH), 7.03 (2H, m, aromatics), 7.41 (2H, m, aromatics).
N-Dimethylcarbamoylmethoxy-2-(2,2-dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-(4-fluorobenzyl)-acetamide was treated with methanol as described in the preparation of Compound 44-D and gave the title ester as white crystals (54% yield); mp 133–135° C. 1HNMR 400 MHz (CDCl3) δ (ppm): 2.9 (3H, s, NCH3), 2.98 (3H, s, NCH3), 3.91 (3H, s, OCH3), 4.54 (2H, s, CH2), 4.96 (2H, s, CH2), 6.52 (1H, s, CH), 7.06 (2H, m, aromatics), 7.39 (2H, m, aromatics), 13.38 (1H, broad s, OH). Anal. calcd for Cl6H19FN2O6: C, 54.24; H, 5.40; N, 7.90. Found: C, 53.62; H, 5.40; N, 7.79.
3-[Dimethylcarbamoylmethoxy-(4-fluorobenzyl)-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 72-B) was treated with paraformaldehyde and methylamine as described in the preparation of Compound 12 to give the title compound as a white solid (25% yield); mp 147–149° C. 1HNMR 400 MHz (CDCl3) δ (ppm): 2.79 (3H, s, NCH3), 2.96 (3H, s, NCH3), 3.11 (3H, s, NCH3), 4.25 (2H, s, CH2), 4.51 (2H, s, CH2), 4.96 (2H, s, CH2), 7.06 (2H, m, aromatics), 7.38 (2H, m, aromatics), 11.54 (1H, broad s, OH). HRMS (ES+) calculated for C17H21FN3O5, [M+H]+: 366.146524. found: 366.146176.
N-[Bis-(4-fluorophenyl)-methyl]-2-(2,2-dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-methoxy-acetamide, prepared using the methods described in the previous examples, was treated in methanol with paraformaldehyde and methylamine as described in the preparation of Compound 44, Method 44B, to give the title compound as a tan solid (42% yield). 1HNMR 400 MHz (DMSO-d6) δ (ppm): 2.9 (3H, broad s, NCH3), 3.1 (3H, broad s, OCH3), 4.2 (2H, broad, NCH2), 6.9 (1H, broad s, CH), 7.0–7.4 (8H, m, aromatics). HRMS (ES+) calculated for C20H22F2N3O4, [M+NH4]+: 406.157838. found: 406.158046.
N-(2-Chloro-4-fluorobenzyl)-2-(2,2-dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-methoxy-acetamide, prepared using the methods described in the previous examples, was treated in methanol with paraformaldehyde and methylamine as described in the preparation of Compound 44, Method 44B, to give the title compound as white crystals (55% yield); mp 152–155° C. 1HNMR 400 MHz (CDCl3) δ (ppm): 3.11 (3H, s, NCH3), 3.75 (3H, s, OCH3), 4.17 (2H, s, NCH2), 5.01 (2H, s, NCH2), 6.99 (1H, m, aromatic), 7.15 (1H, dd, J=2.5 Hz, J=8.0 Hz, aromatic), 7.36 (1H, dd, J=5.7 Hz, J=8.8 Hz, aromatic). Anal. calcd for C14H14ClFN2O4: C, 51.15; H, 4.29; N, 8.52. Found: C, 50.62; H, 4.18; N, 8.40.
N-[1-(4-Chloro-benzyl)-2-(4-chloro-phenyl)-ethyl]-2-(2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-methyl-acetamide, prepared using the methods described in the previous examples, was treated in methanol with paraformaldehyde and methylamine as described in the preparation of Compound 44, Method 44B, to give the title compound as white crystals (56% yield); mp 152–154° C. 1HNMR 400 MHz (DMSO-d6) δ (ppm): mixture of rotamers: 2.7–3.0 (4H, m, CH2), 2.84 (3H, s, NCH3), 2.9 (3H, broad s, NCH3), 3.4 and 3.7 (2H, two broad s, NCH2), 4.16 and 4.9 (1H, broad m, CH), 7.15–7.34 (8H, m, aromatics), 10.7 and 11.0 (1H, two s, OH). HRMS (MAB N2) calculated for C22H22Cl2N2O3: [M]+: 432.100748. found: 432.100835.
4-Flourobenzyl amine (27 grams, 0.22 mole) was dissolved in 200 mL of CH2Cl2. To this was added 400 mL of 1N NaOH and the resulting mixture cooled to 0° C. 4-Fluorobenzoyl chloride (33 grams, 0.21 mole) was added dropwise with stirring. The reaction was allowed to proceed for 20 min after which the organic layer was separated, dried over Na2SO4, filtered and the solvent removed under vacuum to yield 47 grams (92% yield) of 4-fluoro-N-(4-fluoro-benzyl)-benzamide as a solid. MS (M−H) calcd for C14H10F2NO: 246.1. found: 246.0. Anal. Calcd for C14H11F2NO; C, 68.01; H, 4.48; N, 5.66; found: C, 67.76; H, 4.54; N, 5.45. 1H NMR (500 MHz, DMSO) δ: 4.45 (d, 2, J=6), 7.15 (m, 2), 7.34 (overlapping m, 4), 7.97 (m, 2), 9.09 (t, 1, J=6). 13C NMR (125 MHz, DMSO) δ: 41.88, 114.81, 114.98, 115.06, 115.23, 129.08, 129.15, 129.76, 129.83, 130.60, 130.62, 135.66, 135.68, 160.10, 162.03, 162.83, 164.81, 165.04.
4-Fluoro-N-(4-fluoro-benzyl)-benzamide (40.0 grams, 0.16 mole) was dissolved in 240 mL of THF. To this was added BF3.Et2O (7.4 mL. 0.06 mole) and the resulting mixture heated to reflux for 15 min. After cooling to −30° C., BH3.SMe2 (22.3 mL, 0.24 mole) was added using a dropping funnel. The reaction was allowed to warm to room temperature. The reaction flask was fitted with a distillation condenser and solvent removed under reflux for 25 min. The distillation apparatus was replaced with a reflux condenser and the reaction heated to 110° C. for 2 h. After cooling to 0° C., 100 mL of 6N HCl was added and the mixture heated to reflux for 1 hr to yield a thick slurry. 300 mL of 6N NaOH was slowly added at room temperature with intermittent cooling using an ice bath. After all the solid has dissolved Et2O was added and the mixture transferred to a separatory funnel. The organic layer was separated, dried over Na2SO4, filtered and the solvent removed under vacuum to yield an oil. The oil was dissolved in 50 mL of Et2O and 4N HCl (dioxane) added resulting in the formation of a white precipitate which was isolated by filtration to yield 40 grams (93% yield) of bis-(4-fluoro-benzyl)-amine hydrochloride. MS (M+H) calcd for C14H14F2N, 234.1. found: 234.0. Anal. calcd for C14H14F2NCl: C, 62.34; H, 5.23; N, 5.19. found: C, 61.89; H, 5.15; N, 5.27. 1H NMR (500 MHz, DMSO) δ: 4.12 (br s, 4), 7.26 (m, 4), 7.65 (m, 4), 9.91 (br s, 2).
Bis-(4-fluoro-benzyl)-amine hydrochloride (43 grams, 0.16 mole) was suspended in 200 mL of CH2Cl2. To this was added 730 mL of 1N NaOH. The reaction mixture was stirred vigorously while AcCl (20 mL, 0.28 mole) was slowly added. The reaction was stirred for 0.5 h, then the organic layer separated, washed with 1N HCl, dried over Na2SO4, filtered and the solvent removed under vacuum to yield 24 grams (55% yield) of N,N-bis-(4-fluoro-benzyl)-acetamide as an oil. MS (M+H) calcd for C16H16F2NO: 276.1. found: 276.0. Anal. calcd for C16H15F2NO: C, 69.80; H, 5.49; N, 5.08. found: C, 69.53; H, 5.41; N, 5.06. 1H NMR (500 MHz, DMSO) δ: 2.10 (s, 3), 4.45 (s, 2), 4.50 (s, 2), 7.11–7.27 (overlapping m, 8). 13C NMR (125 MHz, DMSO) δ: 21.34, 50.04, 54.80, 114.90, 115.07, 115.27, 115.45, 128.51, 128.58, 129.57, 129.64, 133.32, 133.34, 133.86, 133.89, 160.25, 160.33, 162.27, 170.22.
N,N-Bis-(4-fluoro-benzyl)-acetamide (15.0 grams, 54.5 mmol) and dimethyloxalate (9.6 grams, 81.3 mmol) were dissolved in 54 mL of THF. After cooling to 0° C. 108 mL of 1N LiHMDS (THF) was added dropwise. The reaction mixture was stirred 1 hr then quenched with 1N HCl. The resulting mixture was extracted with CH2Cl2, dried over Na2SO4, filtered and the solvent removed under vacuum. The product was purified by flash column chromatography (SiO2, 80:20 hexanes/EtOAc) to yield 10 grams (53% yield) of 3-[bis-(4-fluoro-benzyl)-carbamoyl]-2-hydroxy-acrylic acid methyl ester as a solid. Mp=118–120° C. Anal. calcd for C19H17F2NO4: C, 63.15; H, 4.74; N, 3.87. found: C, 62.97; H, 4.72; N, 3.81. 1H NMR (500 MHz, CDCl3) δ: 3.87 (s, 3), 4.46 (s, 2), 4.58 (s, 2), 6.32 (s, 1), 7.00–7.26 (overlapping m, 8). 13C NMR (125 MHz, CDCl3) δ: 47.34, 49.35, 53.03, 93.34, 115.66, 115.83, 116.00, 116.18, 128.45, 128.52, 130.03, 130.10, 131.09, 131.93, 131.95, 160.29, 161.44, 161.51, 163.10, 163.41, 163.47, 171.36.
To 2 mL of AcOH at 60° C. was added 1 mL 2M MeNH2 (THF) and 60 mg paraformaldehyde. After stirring for 5 min 3-[bis-(4-fluoro-benzyl)-carbamoyl]-2-hydroxy-acrylic acid methyl ester (723 mg, 2.0 mmol) was added and the resulting solution stirred at 60° C. for 2 h. The reaction mixture was then cooled to room temperature and transferred to a separatory funnel. The solution was extracted with EtOAc, the organic layer separated, washed with H2O, satd NaCl, then dried over Na2SO4. After filtration the solvent was removed to isolate 500 mg (67% yield) of 4-hydroxy-1-methyl-5-oxo-2,5-dihydro-1H-pyrrole-3-carboxylic acid bis-(4-fluoro-benzyl)-amide as an orange solid. MS (M+H) calcd for C20H19F2N2O3: 373.1. found: 373.0. 1H NMR (500 MHz, DMSO) δ: 2.94 (s, 3), 4.01 (s, 2), 4.46 (br s, 2), 4.55 (br s, 2), 7.13–7.22 (overlapping m, 8).
From N-(4-chlorobenzyl)-3,4-dichlorobenzamide (Borgma et al. Farmaco Ed. Sci. 1977, 32, 813). 1HNMR 400 MHz (C6D6) δ (ppm): 3.22 (2H, s, NCH2), 3.31 (2H, s, NCH2), 6.78 (1H, d, J=8.5 Hz, aromatic), 6.99 (2H, d, J=8.1 Hz, aromatics), 7.15 (2H, d, J=8.1 Hz, aromatics), 7.24–7.31 (3H, m, aromatics).
4-(Chlorobenzyl)-(3,4-dichlorobenzyl)-amine was acetylated as described in the preparation of Compound 76-C and gave the title amide as a clear oil (78% yield). 1HNMR 400 MHz (C6D6) δ (ppm), mixture of rotamers: 1.77 and 1.78 (3H, 2 s, COCH3), 3.6 and 3.66 (2H, 2 s, NCH2), 4.26. and 4.33 (2H, 2 s, NCH2), 6.37–7.2 (7H, m, aromatics). Anal. calcd for C16H14Cl3NO: C, 56.08; H, 4.12; N, 4.09. Found: C, 56.13; H, 4.07; N, 4.08.
N-(4-Chlorobenzyl)-N-(3,4-dichlorobenzyl)-acetamide was reacted with dimethyl oxalate as described in the preparation of Compound 76-D and gave the title methyl ester as a clear oil (40% yield). 1HNMR 400 MHz (CDCl3) δ (ppm): mixture of rotamers: 3.9 (3H, s, OCH3), 4.47, 4.5, 4.58 and 4.62 (2×2H, 4 s, NCH2), 6.27 and 6.33 (1H, 2 s, CH), 7.0–7.48 (7H, m, aromatics), 14.33 and 14.37 (1H, 2 s, OH).
3-[(4-Chlorobenzyl)-(3,4-dichlorobenzyl)-carbamoyl]-2-hydroxy-acrylic acid methyl ester was reacted with paraformaldehyde and methylamine as described in the preparation of Compound 12 to give the title compound as a white solid (39% yield); mp 165° C., dec. 1HNMR 400 MHz (CDCl3) δ (ppm): 3.09 (3H, s, NCH3), 4.09, 4.55 and 4.56 (3×2H, 3 s, NCH2), 7.05–7.45 (7H, m, aromatics), 10.3 (1H, broad, OH).
3,4-(Dichlorophenyl)-(4-fluorobenzyl)-amine was acetylated as described in the preparation of Compound 76-C and gave the title amide as a clear oil (89% yield). 1HNMR 400 MHz (C6D6) δ (ppm): 1.59 (3H, s, COCH3), 4.56 (2H, s, NCH2), 6.04 (1H, broad s, aromatic), 6.71–6.98 (6H, m, aromatics). Anal. calcd for C15H12Cl2FNO: C, 57.71; H, 3.87; N, 4.48. Found: C, 57.85; H, 3.89; N, 4.49.
N-(3,4-Dichlorophenyl)-N-(4-fluorobenzyl)-acetamide was reacted with dimethyl oxalate as described in the preparation of Compound 76-D and gave the title methyl ester as a clear oil (40% yield). 1HNMR 400 MHz (CDCl3) δ (ppm): 3.84 (3H, s, OCH3), 4.90 (2H, s, NCH2), 5.69 (1H, s, CH), 6.83 (1H, dd, J=2.0 Hz and J=8.5 Hz, aromatic), 7.02 (2H, m, aromatic), 7.19 (3H, m, aromatic), 7.47 (1H, d, J=8.5 Hz, aromatic), 13.83 (1H, s, OH).
3-[(3,4-Dichlorophenyl)-(4-fluorobenzyl)-carbamoyl]-2-hydroxy-acrylic acid methyl ester (Compound 78-B) was treated with paraformaldehyde and methylamine as described in the preparation of Compound 12 to give the title compound as a white solid (68% yield); mp 195° C., dec. 1HNMR 400 MHz (CDCl3) δ (ppm): 2.9 (3H, s, NCH3), 3.03 (2H, s, NCH2), 4.90 (2H, s, NCH2), 6.87 (1H, dd, J=2.5 Hz and J=8.6 Hz, aromatic), 7.02 (2H, m, aromatics), 7.2 (2H, m, aromatics), 7.22 (1H, d, J=2.5 Hz, aromatic), 7.49 (1H, d, J=8.6 Hz, aromatic), 11.9 (1H, broad, OH). Anal. calcd for C19H15Cl2FN2O3: C, 55.76; H, 3.69; N, 6.84. Found: C, 55.53; H, 3.61; N, 6.75.
Compound 79-A was prepared using methods described in the previous examples. 1H NMR and 13C NMR show a mixture of rotamers at room temperature. 1H NMR (500 MHz, CDCl3) δ: 2.20 (s), 2.95 (s), 2.97 (s), 4.56 (s), 4.62 (s), 7.19–7.41 (overlapping m). 13C NMR (125 MHz, CDCl3) δ: 21.42, 21.80, 33.82, 35.58, 50.68, 54.29, 126.33, 127.41, 127.70, 128.07, 128.63, 128.99, 136.46, 137.26, 170.88, 171.19.
Compound 79-B was prepared using methods described in the previous examples. 1H NMR shows a mixture of rotamers at room temperature. 1H NMR (500 MHz, CDCl3) δ: 3.02 (s), 3.86 (s), 3.90 (s), 4.60 (s), 4.66 (s), 6.31 (s), 6.34 (s), 7.18–7.40 (overlapping m).
Compound 79 was prepared using methods described in the previous examples. HRMS (M−H) calcd for C14H15N2O3: 259.10827. found 259.1082. 1H NMR (500 MHz, CDCl3) δ: 3.03 (s, 3), 3.09 (s, 3), 4.15 (s, 2), 4.65 (s, 2), 7.23–7.37 (m, 5). 13C NMR (125 MHz, CDCl3) δ: 29.95, 34.67, 50.80, 52.28, 108.72, 127.38, 127.86, 128.94, 136.12, 154.15, 165.10, 166.46.
PL-FMP resin (4-formyl-3-methoxy-phenoxymethyl functionalized polystyrene), VII-1 in Scheme VII or similar aldehyde derivatized resin (approximately 40 mg, 0.048 meq.) was suspended in 2 ml of anhydrous DMF, anhydrous trimethylorthoformate and acetic acid mixture in 7:3:1 ratio. To this mixture was added a primary amine, VII-2 in scheme VII, (0.72 mmol) followed by sodium triacetoxy borohydride (0.72 mmol). The resulting mixture was agitated for 72 h at room temperature using a shaker. The resin was filtered, washed successively with (DMF, THF and DCM, 3×2 ml each), dried and used as is in the next step.
The resin (VII-3 in Scheme VII) was suspended in anhydrous dichloromethane (1 mL) and to this was added, 2,2-dimethyl-5-(carboxymethylene)-1,3-dioxalan-4-one (Compound III-A) (16.5 mg, 0.096 mmol), Pybop (50 mg, 0.096 mmol) and iPr2NEt (24.8 mg, 0.192 mmol). The resulting mixture was agitated for 48 h at room temperature using a shaker. Resin was filtered, washed successively (DMF, THF and DCM, 3×2 ml each), dried and used in the next step.
Next the resin, VII-4 in Scheme VII, was suspended in 1 ml anhydrous N-methyl-2-pyrrolidinone (NMP). To this mixture was added a preformed imine mixture (preformed by heating aldehyde (18 mg, 0.55 mmol), and amine (0.5 mmol) and 0.25 ml methanol, at 70° C. for 2 h) and the resulting mixture was heated with agitation in a sealed container at 70–80° C. for 72 h. Resin was filtered, washed successively with (DMF, DMF/MeOH (1:1), THF and DCM, 3×2 ml each), dried.
The final product, VII-6 in Scheme VII, was cleaved from the resin by treating with 1.5 ml of 1:1 mixture of trifluoroacetic acid and dichloromethane for 1 h. TFA solution was filtered and the solvent was evaporated to give the required product.
Compounds 81-A to 81-V were synthesized from [[2-(2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl]-(4-fluorobenzyl)-aminooxy]-acetic acid according to Scheme VIII. Amines VIII-3 (0.165 mmol) in 1,2-dichloroethane (1 ml) were treated at 5° C. with 2-(2-pyridyl)ethyl functionalized silica gel (0.38 mmol) followed by a solution of [[2-(2,2-dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-acetyl]-(4-fluorobenzyl)-aminooxy]-acetyl chloride, VIII-2, (0.165 mmol) in 1,2-dichloroethane (1 ml). After one hour at 25° C., the reaction mixtures were filtered and purified on a Shimadzu automated preparative HPLC system (column Waters X Terra C-8, 5μ, 19×100 mm, elution H2O5 mM NH4Oac-acetonitrile). The collected compounds were analysed using the following LC/MS conditions.
Spectrometry (MS) data were determined with a Micromass ZMD Platform TSQ 7000 LC/MS in positive electrospray mode.
According to the method illustrated in Scheme VIII, the dioxolanes 81-A to 81-V, VIII-4 in the scheme, (approximately 0.06 mmol) were treated with a solution of formaldehyde-methyl amine adduct (0.12 mmol) in methanol (1 ml) as described in the preparation of Compound 44 (Method 44B) and the resulting mixtures were heated at 50° C. for 45 min. The reaction mixture was then diluted with acetonitrile (1 ml) and purified on a Shimadzu automated preparative HPLC system (column Waters X Terra C-8, 5 μ, 19×100 mm, elution water 0.05% TFA-acetonitrile). The collected compounds were analysed using the following LC/MS conditions.
Spectrometry (MS) data were determined with a Micromass ZMD Platform TSQ 7000 LC/MS in positive electrospray mode.
The general method for the synthesis of compounds 747–750 is outlined in Scheme X.
To a solution of 2-aminoacetaldehyde dimethylacetal (0.526 g mL, 5.0 mmol) and paraformaldehyde 90.15 g, 5 mmol) in MeOH (10 mL) at 55° C. was added N-(3,4-dichloro-benzyl)-2-(2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-methyl-acetamide (1.715 g, 5.0 mmol). After 45 min, the reaction mixture was cooled and purified by preparative HPLC on a C18 reverse phase column using acetonitrile (40–50%)/water (0.1% TFA) as eluent. The fractions containing the desired product were combined, concentrated and lyophilized to yield the title compound as an amber oil (0.68 g, 34% yield). 1H NMR (300 MHz, CDCl3) δ: 10.81 (bs, 1H), 7.42 (d, 1H, J=8.1), 7.35 (d, 1H, J=1.8), 7.10 (dd, 1H, J=8.1, J=1.8), 4.59 (s, 2H), 4.47 (t, 1H, J=5.1), 4.27 (s, 2H), 3.59 (d, 2H, J=5.1), 3.38 (s, 6H), 3.02 (s, 3H). HRMS (M+Na) calcd for C17H20N2Cl2O5Na: 425.0647. found: 425.0647.
A solution of 1-(2,2-dimethoxy-ethyl)-4-hydroxy-5-oxo-2,5-dihydro-1H-pyrrole-3-carboxylic acid (3,4-dichloro-benzyl)-methyl-amide (0.29 g, 0.71 mmol) in acetonitrile/water (10 mL, 2.5:1) was stirred with trifluoroacetic acid (1 mL) overnight at room temp. The mixture was concentrated then dissolved in MeOH (3 mL). 1-(4-pyridyl)-piperazine (0.42 g, 2.6 mmol) was added and the resulting mixture was stirred at room temp 30 min. Sodium cyanoborohydride (0.013 g, 0.2 mmol) was added and the mixture was stirred an additional 5 h at room temp. The crude product was purified by preparative HPLC (YMC Combiprep ODS-A, 30 mm×50 mm, MeOH/H2O/0.1% TFA) to yield the title compound as an amber foam (0.0042 g, 12% yield). HRMS (M+H) calcd for C24H28N5Cl2O3: 504.15693; found: 504.1564.
Compound 748 was prepared from 1-(2,2-dimethoxy-ethyl)-4-hydroxy-5-oxo-2,5-dihydro-1H-pyrrole-3-carboxylic acid (3,4-dichloro-benzyl)-methyl-amide and 1-(4-fluoro-phenyl)-piperazine using the method described for compound 747. The title compound was isolated as an amber foam (0.009 g, 25% yield). HRMS (M+H) calcd for C24H28N4Cl2O3F: 521.15226. found: 521.1531.
Compound 749 was prepared from 1-(2,2-dimethoxy-ethyl)-4-hydroxy-5-oxo-2,5-dihydro-1H-pyrrole-3-carboxylic acid (3,4-dichloro-benzyl)-methyl-amide and 1-benzo[1,3]dioxol-5-ylmethyl-piperazine using the method described for compound 747. The title compound was isolated as a white solid (0.0068 g, 17% yield). HRMS (M+H) calcd for C27H31N4Cl2O5: 561.16716. found: 561.1674.
Compound 750 was prepared from 1-(2,2-dimethoxy-ethyl)-4-hydroxy-5-oxo-2,5-dihydro-1H-pyrrole-3-carboxylic acid (3,4-dichloro-benzyl)-methyl-amide and 2,6-dimethyl-morpholine using the method described for compound 747. The title compound was isolated as a pale yellow powder (0.014 g, 14% yield). HRMS (M+H) calcd for C21H28N3Cl2O4: 456.14570. found: 456.1472.
The general method for the synthesis of compounds 751–758 is outlined in Scheme XI.
To a solution of 4-{4-[(3,4-dichloro-benzyl)-methyl-carbamoyl]-3-hydroxy-2-oxo-2,5-dihydro-pyrrol-1-yl}-butyric acid, Compound 24, (0.06 g, 0.15 mmol) in dichloromethane (1 mL) was added EDC (0.057 g, 0.3 mmol), HOBT (0.0020 g, 0.015 mmol) and dimethyl amine (0.15 mL, 2 M solution in THF, 0.30 mmol). The mixture was stirred at room temp for 6 h then concentrated and the crude product purified by preparative HPLC (YMC Combiprep ODS-A, 30 mm×50 mm, MeOH/H2O/0.1% TFA) to yield the title compound as an amber oil (0.0147 g, 23% yield). HRMS (M+H) calcd for C19H24N3Cl2O4: 428.11440. found: 428.1143.
Compound 752 was prepared from 4-{4-[(3,4-dichloro-benzyl)-methyl-carbamoyl]-3-hydroxy-2-oxo-2,5-dihydro-pyrrol-1-yl}-butyric acid and morpholine using the method described for compound 751. The title compound was isolated as an amber oil (0.0148 g, 21% yield). HRMS (M+H) calcd for C21H26N3Cl2O5: 470.12496. found: 470.1256.
Compound 753 was prepared from 4-{4-[(3,4-dichloro-benzyl)-methyl-carbamoyl]-3-hydroxy-2-oxo-2,5-dihydro-pyrrol-1-yl}-butyric acid and 4-methyl-piperazine using the method described for compound 751. The title compound was isolated as a white powder (0.0508 g, 42% yield). HRMS (M+H) calcd for C22H29N4Cl2O4: 483.1566. found: 483.1581.
Compound 754 was prepared from 4-{4-[(3,4-dichloro-benzyl)-methyl-carbamoyl]-3-hydroxy-2-oxo-2,5-dihydro-pyrrol-1-yl}-butyric acid and methylamine using the method described for compound 751. The title compound was isolated as a white powder (0.0192 g, 18% yield). HRMS (M+H) calcd for C18H22N3Cl2O4: 414.09875. found: 414.0969.
Compound 755 was prepared from 4-{4-[(3,4-dichloro-benzyl)-methyl-carbamoyl]-3-hydroxy-2-oxo-2,5-dihydro-pyrrol-1-yl}-butyric acid and methane sulfonamide using the method described for compound 751. The title compound was purified by preparative HPLC (C18, ODS-A, S-75 μm, 30%–40% acetonitrile/water/0.5% TFA) and isolated as a white powder (0.0158 g, 13% yield). HRMS (M−H) calcd for C18H20N3Cl2O6S: 476.04499; found: 476.0431.
Compound 756 was prepared from 4-{4-[(3,4-dichloro-benzyl)-methyl-carbamoyl]-3-hydroxy-2-oxo-2,5-dihydro-pyrrol-1-yl}-butyric acid and 3,5-dimethyl-isoxazole-4-sulfonic acid amide using the method described for compound 751. The title compound was purified by preparative HPLC (C18, ODS-A, S-75 μm, 40% acetonitrile/water/0.5% TFA) and isolated as a white powder (0.0296 g, 21% yield). HRMS (M−H) calcd for C22H23N4Cl2O7S: 557.06645. found: 557.0663.
Compound 757 was prepared from 4-{4-[(3,4-dichloro-benzyl)-methyl-carbamoyl]-3-hydroxy-2-oxo-2,5-dihydro-pyrrol-1-yl}-butyric acid and 1-phenyl-cyclopropanesulfonic acid amide using the method described for compound 751. The title compound was purified by preparative HPLC (C18, ODS-A, S-75 μm, 50% acetonitrile/water/0.5% TFA) and isolated as a white powder (0.0026 g, 3% yield). HRMS (M−H) calcd for C26H30N3Cl2O6S: 582.12324. found: 582.1215.
Compound 758 was prepared from {4-[(3,4-dichloro-benzyl)-methyl-carbamoyl]-3-hydroxy-2-oxo-2,5-dihydro-pyrrol-1-yl}-acetic acid, Compound 20, and morpholine using the method described for compound 751. The title compound was isolated as a white powder (0.0088 g, 20% yield). HRMS (M+H) calcd for C19H22N3Cl2O5: 442.09366. found: 442.0951.
The general method for the synthesis of compounds 759–765 is outlined in Scheme XII.
Compound 84-A was prepared from N-(3,4-dichloro-benzyl)-2-(2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-methyl-acetamide and 2-piperazin-1-yl-ethylamine using the method described for compound 37. The title compound was isolated as a white solid (0.183 g, 86% yield). HRMS (M+H) calcd for C19H25N3Cl2O4: 427.13038. found: 427.1307.
To a solution of benzoyl chloride (0.023 mL, 0.23 mmol) in dichloromethane (1 mL) cooled to 0° C. was added dropwise a solution of 4-hydroxy-5-oxo-1-(2-piperazin-1-yl-ethyl)-2,5-dihydro-1H-pyrrole-3-carboxylic acid (3,4-dichloro-benzyl)-methyl-amide (0.10 g, 0.20 mmol) in pyridine/dichloromethane (4 mL, 1:1). The resulting mixture was stirred at room temp 18 h and quenched with 1N HCl. The organic phase was washed with 1N HCl (3 times). The aqueous washings were combined and purified by preparative HPLC (C18, ODS-A, S-75 μm, 30% acetonitrile/water/0.5% TFA) to give the title compound as a white powder (0.0215 g, 20% yield). HRMS (M+H) calcd for C26H29N4Cl2O4: 531.1566; found: 531.1563.
Compound 760 was prepared from 4-hydroxy-5-oxo-1-(2-piperazin-1-yl-ethyl)-2,5-dihydro-1H-pyrrole-3-carboxylic acid (3,4-dichloro-benzyl)-methyl-amide and 4-fluoro-benzoyl chloride using the method described for compound 759. The title compound was purified by preparative HPLC (C18, ODS-A, S-75 μm, 30% acetonitrile/water/0.5% TFA) and isolated as a white powder (0.0098 g, 2% yield). HRMS (M−H) calcd for C26H26N4Cl2O4F: 547.13151. found: 547.1310.
Compound 761 was prepared from 4-hydroxy-5-oxo-1-(2-piperazin-1-yl-ethyl)-2,5-dihydro-1H-pyrrole-3-carboxylic acid (3,4-dichloro-benzyl)-methyl-amide and 4-methyl-benzoyl chloride using the method described for compound 759. The title compound was purified by preparative HPLC (C18, ODS-A, S-75 μm, 30%–40% acetonitrile/water/0.5% TFA) and isolated as a white powder (0.0210 g, 17% yield). HRMS (M+H) calcd for C27H31N4Cl2O4: 545.17225. found: 545.1720.
Compound 762 was prepared from 4-hydroxy-5-oxo-1-(2-piperazin-1-yl-ethyl)-2,5-dihydro-1H-pyrrole-3-carboxylic acid (3,4-dichloro-benzyl)-methyl-amide and pyridine-2-carbonyl chloride using the method described for compound 759. The title compound was purified by preparative HPLC (C18, ODS-A, S-75 μm, 10%-20%-30% acetonitrile/water/0.5% HCl) and isolated as a brown solid (0.0476 g, 39% yield). HRMS (M+H) calcd for C25H28N4Cl2O5: 532.15184. found: 532.1514.
Compound 763 was prepared from 4-hydroxy-5-oxo-1-(2-piperazin-1-yl-ethyl)-2,5-dihydro-1H-pyrrole-3-carboxylic acid (3,4-dichloro-benzyl)-methyl-amide and isoxazole-5-carbonyl chloride using the method described for compound 759. The title compound was purified by preparative HPLC (C18, ODS-A, S-75 μm, 10%–20%–30% acetonitrile/water/0.5% HCl) and isolated as a brown solid (0.0268 g, 22% yield). HRMS (M+H) calcd for C23H26N5Cl2O5: 522.13111. found: 522.1312.
Compound 764 was prepared from 4-hydroxy-5-oxo-1-(2-piperazin-1-yl-ethyl)-2,5-dihydro-1H-pyrrole-3-carboxylic acid (3,4-dichloro-benzyl)-methyl-amide and methanesulfonyl chloride using the method described for compound 759. The title compound was purified by preparative HPLC (C18, ODS-A, S-75 μm, 10%–20%–30% acetonitrile/water/0.5% HCl) and isolated as a brown solid (0.0140 g, 12% yield). HRMS (M+H) calcd for C20H27N4Cl2O5S: 505.10793. found: 505.1095.
Compound 765 was prepared from 4-hydroxy-5-oxo-1-(2-piperazin-1-yl-ethyl)-2,5-dihydro-1H-pyrrole-3-carboxylic acid (3,4-dichloro-benzyl)-methyl-amide and dimethylsulfamoyl chloride using the method described for compound 759. The title compound was purified by preparative HPLC (C18, ODS-A, S-75 μm, 30% acetonitrile/water/0.5% HCl) and isolated as a brown solid (0.0231 g, 19% yield). HRMS (M+H) calcd for C21H30N5Cl2O5S: 534.13448. found: 534.1322.
To a solution of 4-hydroxy-1-(2-morpholin-4-yl-ethyl)-5-oxo-2,5-dihydro-1H-pyrrole-3-carboxylic acid (3,4-dichloro-benzyl)-methyl-amide (0.069 g, 0.16 mmol) in dichloromethane (1 mL), was added meta-chloro-peroxy-benzoic acid (MCPBA) (0.036 g, 0.21 mmol). The mixture was stirred for 24 h then concentrated. Purification using preparative HPLC (C18, ODS-A, S-75 μm, 30% acetonitrile/water/0.5% HCl) gave the title compound as a white solid (0.03 g, 94% yield). HRMS (M+H) calcd for C19H24N3Cl2O5: 444.10931. found: 444.1073.
The general method for the preparation of compounds 767–777 is outlined in Scheme XIII.
2-Aminoethanol (0.061 g, 1.0 mmol) and paraformaldehyde (0.03 g, 1.0 mmol) were stirred in methanol (10 mL) at 55° C. until the solids dissolved (approximately 20 min). The solution was cooled and used without further purification
To a solution of 2-methyleneamino-ethanol (1 mL, 0.1 mmol) in MeOH (1 mL) was added N-(3,4-dichloro-benzyl)-2-(2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-methyl-acetamide (0.036 g, 0.1 mmol). The mixture was stirred at 55° C. for 1 h, cooled and purified by preparative HPLC (YMC Combiprep ODS-A, 30 mm×50 mm, MeOH/H2O/0.1% TFA). The title compound was isolated as a white foam (0.018 g, 49% yield). HRMS (M−H) calcd for C15H15N2Cl2O4: 357.04089. found: 357.0396.
Compound 768 was prepared from N-(3,4-dichloro-benzyl)-2-(2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-methyl-acetamide, paraformaldehyde and 2-amino-3-(1H-indol-3-yl)-propan-1-ol according to the procedures described for compound 86-A and compound 767. The title compound was isolated as a yellow solid (0.0038 g, 8% yield). 1HNMR (300 MHz, CDCl3) δ: 7.52 (1H, d, J=8.05 Hz), 7.36–7.29 (3H, m), 7.14–6.98 (4H, m), 4.46 (2H, s), 4.40–4.36 (1H, m), 3.99 (2H, s), 3.81–3.78 (2H, m), 3.11–3.08 (3H, m), 2.85 (3H, s).
As illustrated in Scheme XII, compounds 769–777 were prepared from N-(3,4-chloro-benzyl)-2-(2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-methyl-acetamide, paraformaldehyde and an amine, I-7, according to the procedures described for compound 86-A and compound 767.
Reaction of (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl chloride with methyl-naphthalen-1-yl-methyl-amine as described in the preparation of Compound 44-C gave the title amide as a, pale yellow oil (0.4931 g, 26% yield). 1HNMR (300 MHz, CDCl3) δ: 8.18 (0.6H, d), 7.88–7.80 (2.4H, m), 7.54–7.38 (4H, m), 6.17 (0.6H, s), 6.08 (0.4H, s), 5.13 (1.2H, s), 5.06 (0.8H, s), 3.08 (1.2H, s), 2.90 (1.8H, s), 1.73 (3.6H, s), 1.67 (2.4H, s).
2-(2,2-Dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-methyl-N-naphthalen-1-ylmethyl-acetamide was treated with methanol as described in the preparation of Compound 44-D and gave the title ester as a colorless oil (0.04 g, 33% yield). 1HNMR (300 MHz, CDCl3) δ: 14.65 (1H, bs), 7.89–7.82 (3H, m), 7.57–7.20 (4H, m), 6.31 (0.6H, s), 6.21, (0.4H, s), 5.15 (1.2H, s), 5.07 (0.8H, s), 3.89 (1.8H, s), 3.80 (1.2H, s), 3.11 (1.2H, s), 2.94 (1.8H, s).
2-Hydroxy-3-(methyl-naphthalen-1-ylmethyl-carbamoyl)-acrylic acid methyl ester was treated with paraformaldehyde and an amine according to the procedure described in the preparation of compound 12 to yield compounds 778–781.
2-Hydroxy-3-(methyl-naphthalen-1-ylmethyl-carbamoyl)-acrylic acid methyl ester was treated with paraformaldehyde and 4-amino-butyric acid according to the procedure described in the preparation of compound 12 to give the title compound as a white powder (0.0210 g, 55% yield). 1HNMR (300 MHz, CDCl3) δ: 7.93–7.81 (2H, m), 7.55–7.42 (4H, m), 7.42–7.28 (1H, m), 5.13 (2H, s), 4.14 (2H, s), 3.52–3.50 (2H, m), 2.36–2.29 (2H, m), 1.90–1.87 (2H, m).
The general method for the preparation of compounds 783–791 is illustrated in Scheme XV.
A suspension of 2,4-difluorobenzaldehyde (16 mL, 146 mmol) and sodium thiomethoxide (14 g, 200 mmol) in toluene (200 mL) was stirred at 80° C. for 7 h and 14 h at room temperature. The reaction mixture was diluted with ether (300 mL), washed with water (100 mL), saturated aqueous NaHCO3 (100 mL) and brine (50 mL). The aqueous layers were combined and extracted with ether (2×100 mL). The organic layers were combined and dried over anhydrous Na2SO4, filtered and concentrated to give a viscous oil. This viscous oil was dissolved in ether/hexanes (1:1, v/v) and slowly concentrated on under vacuum. The precipitated white solid was separated by filteration and dried to give 2-methylthio-4-fluorobenzaldehyde (18.7 g, 75% yield). 1HNMR (500 MHz, CDCl3) δ: 10.11 (1H, s), 7.78 (1H, dd, J=8.55, 6.11 Hz), 6.97 (1H, dd, J=10.07, 2.44 Hz), 6.91 (1H, td, J=8.54, 2.44 Hz), 2.45 (3H, s). MS calcd for C8H7FOS (M+H): 171.2. found: 171.6.
A solution of 2-methylthio-4-fluorobenzaldehyde (2.04 g, 12 mmol) and 2M methylamine in methanol (24 mL, 48 mmol) was stirred at room temperature for 2 h. To this was added a solution of ZnCl2 (0.818 g, 6 mmol) and NaCNBH3 (0.754 g, 12 mmol) in methanol (30 mL). After stirring for 20 h, the reaction mixture was concentrated and the resulting residue was taken up in aqueous NaOH (0.5 M, 20 mL), extracted with CH2Cl2 (5×50 mL). The combined organic layers were concentrated and the resulting residue was taken up in 1N HCl (25 mL). This solution was extracted with ethyl acetate (3×25 mL). The organic layers were discarded and aqueous layer was brought to pH 9 by adding Na2CO3 and extracted with CH2Cl2 (3×50 mL). The combined CH2Cl2 layers were dried over MgSO4, filtered and concentrated to give the desired benzylamine as a viscous pale yellow oil (1.4 g, 60% pure).
To a stirred solution of (4-fluoro-2-methylsulfanyl-benzyl)-methyl-amine (1.3 g) and diisopropylethylamine (1.74 mL, 10 mmol) in CH2Cl2 (60 mL) was added (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl chloride (1.1 g, 6 mmol). After 1 h, the reaction mixture was concentrated and the resulting residue was taken up in ether (100 mL), washed with 1N HCl (10 mL) and brine (10 mL). The organic layer was dried over anhydrous Na2SO4, filtered and concentrated to give a viscous yellow oil. Flash chromatography on a silica gel column with 3:2 hexanes/EtOAc followed by 2:3 hexanes/EtOAc gave the desired product as a 1:1 mixture of E and Z isomers 1.8123 g, 89% yield). 1HNMR (500 MHz, CDCl3) δ: 7.17 (0.5H, dd, J=8.54, 6.11 Hz), 7.00 (0.5H, dd, J=8.55, 5.8 Hz), 6.94–6.89 (1H, m), 6.83–6.77 (1H, m), 6.17 (0.5H, s), 6.04 (0.5H, s), 4.68 (1H, s), 4.51 (1H, s), 2.98 (1.5H, s), 2.97 (1.5H, s), 2.48 (1.5H, s), 2.45 (1.5H, s), 1.72 (3H, s), 1.67 (3H, s). MS calcd for C16H19FNO4S (M+H): 340.1. found: 340.3.
To a stirred solution of 4-fluoro-2-methylsulfanylbenzaldehyde (9.7 g, 57 mmol) in CH2Cl2 (300 mL) was added in small portions m-chloroperbenzoic acid (60%, 20.71 g, 120 mmol) over 20 minutes. After 24 h, mL of dimethylsulfoxide followed by saturated NaHCO3 (100 mL) was added and stirred for additional 2 h. The organic layer separated and washed with saturated NaHCO3 (2×100 mL). The combined aqueous layers were saturated with NaCl and extracted with CH2Cl2 (2×100 mL). The combined organic phases were dried over anhydrous Na2SO4, filtered and concentrated to give a viscous yellow oil. This residue was purified by flash chromatography on a silica gel column using hexanes/ether/ethyl acetate (gradient elution). The fractions containing the desired product are combined and concentrated to give 4-fluoro-2-methylsulfonylbenzaldehyde (3.85 g, 33%) and 4-fluoro-2-methylsulfinylbenzaldehyde (5.61 g, 53% yield) as a white solid. 4-fluoro-2-methylsulfonylbenzaldehyde: 1HNMR (500 MHz, CDCl3) δ: 10.41 (1H, s), 8.08–8.06 (1H, M), 7.85–7.71 (2H, m), 3.09 (3H, s). HRMS calcd for C8H6FO3S (M−H): 201.0022. found: 201.0025. 4-fluoro-2-methylsulfinyl-benzaldehyde: 1HNMR (500 MHz, CDCl3) δ: 9.97 (1H, s), 8.07–8.05 (1H, M), 7.99–7.94 (1H, m), 7.36–7.33 (1H, m), 2.80 (3H, s). MS calcd for C8H8FO2S (M+H): 187.02. found: 187.02.
To a stirred solution of 4-fluoro-2-methylsulfinylbenzaldehyde (2.234 g, 12 mmol) in 2M methylamine in methanol (24 mL, 48 mmol) was added a solution of ZnCl2 (0.818 g, 6 mmol) and NaCNBH3 (0.754 g, 12 mmol) in methanol (30 mL). After stirring for 16 h an additional methyl amine (1 equiv) and NaCNBH3 (1 equiv) every 24 h for 5 days. After seven days, the reaction mixture was concentrated and the resulting residue was taken up into aqueous NaOH (1 M, 100 mL), extracted with CH2Cl2 (5×50 mL). The combined organic layers were concentrated to give a yellow oil which was a mixture of aldehyde and the desired (4-Fluoro-2-methanesulfinyl-benzyl)-methyl-amine. This material was used in the next step without further purification.
To a stirred solution of above amine and diisopropylethylamine (1.75 mL, 10 mmol) in CH2Cl2 (30 mL) was added (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl chloride (0.95 g, 5 mmol). After 1 h, the reaction mixture was concentrated and the resulting residue was taken up into ether (150 mL), washed with 1N HCl (10 mL), water (20 mL) and brine (20 mL). The organic layer was dried over anhydrous Na2SO4, filtered and concentrated to give a yellow residue which was purified by flash column chromatography on silica gel column using 1:1 v/v hexanes/ethyl acetate, ethyl acetate and 2–5% methanol/ethyl acetate. The fractions containing the compound were combined and concentrated to give the product (1.65 g, 93% yield based on acid chloride used). 1HNMR (500 MHz, CDCl3) δ: 7.76–7.72 (1H, m), 7.28 (1H,dd, J=8.3, 5.0 Hz), 7.13 (1H, td, J=8.2, 2.7 Hz), 6.14 (1H, s), 4.88 (1H, d, J=15.3 Hz), 4.46 (1H, d, J=15.3 Hz), 3.03 (3H, s), 2.71 (3H, s), 1.72 (6H, s). LRMS calcd for C16H19FNO5S (M+H): 356.1. found: 356.3.
To a solution of 2-(2,2-Dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-(4-fluoro-2-methylsulfanyl-benzyl)-N-methyl-acetamide (0.5 g, 1.475 mmol) in CH2Cl2 (10 mL) was added 50% m-chloroperbenzoic acid (1.035 g, 3 mmol) and the resulting mixture stirred at room temperature for 4 h, then, taken up into EtOAc, washed successively with saturated NaHSO4, saturated NaHCO3 and brine. The organic layer was dried over anhydrous Na2SO4, filtered, concentrated and the residue purified on a silica gel column using hexanes/EtOAc (0–75%) to afford the desired product as a mixture of E/Z isomers (0.4977 g, 91%). 1HNMR (300 MHz, CDCl3) δ: 7.76 (1H, dd, J=8.2, 2.7 Hz), 7.38–7.27 (2H, m), 6.21 (0.8H, s), 5.93 (0.2H, s), 5.05 (1.6H, s), 5.01 (0.4H, s), 3.19 (2.4H, s), 3.16 (2.4H, s), 3.11 (0.6H, s), 3.03 (0.6H, s), 1.72 (4.8H, s), 1.69 (1.2H, s). LRMS calcd for C16H19FNO6S (M+H): 372. found: 372.
2-(2,2-Dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-(4-fluoro-2-methylsulfanyl-benzyl)-N-methyl-acetamide was reacted with paraformaldehyde and 4-amino-butyric acid as described in Method 44B to give the title compound as a white solid (0.05 g, 49% yield). 1HNMR (300 MHz, CDCl3) δ: 7.11 (1H, dd, J=8.42, 5.85 Hz), 6.91 (1H, dd, J=9.52, 2.57 Hz), 6.82 (1H, td, J=8.41, 2.56 Hz), 4.64 (2H, s), 4.15 (2H, s), 3.55 (2H, t, J=6.96 Hz), 2.98 (3H, s), 2.47 (3H, s), 2.38 (2H, t, J=6.95 Hz), 1.96–1.88 (2H, m).
2-(2,2-Dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-(4-fluoro-2-methanesulfinyl-benzyl)-N-methyl-acetamide was reacted with paraformaldehyde and 4-amino-butyric acid as described in Method 44B to give the title compound as a colorless oil (0.015 g, 18% yield). 1HNMR (300 MHz, CDCl3) δ: 7.76 (1H, dd, J=8.05, 2.20 Hz), 7.34 (1H, dd, J=8.41, 5.12 Hz), 7.19 (1H, td, J=8.05, 2.56 Hz), 4.85 (2H, d, J=15.01 Hz), 4.18 (2H, s), 3.58 (2H, t, J=5.86), 3.01 (3H, s), 2.76 (3H, s), 2.39 (2H, t, J=5.86 Hz), 1.95 (2H, t, J=6.22 Hz).
2-(2,2-Dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-(4-fluoro-2-methanesulfonyl-benzyl)-N-methyl-acetamide was reacted with paraformaldehyde and 4-amino-butyric acid as described in Method 44B to give the title compound as a colorless oil (0.01 g, 9% yield). 1HNMR (300 MHz, CDCl3) δ: 7.78 (1H, dd, J=8.06, 2.57 Hz), 7.43 (1H, dd, J=8.60, 4.94 Hz), 7.36–7.30 (1H, m), 5.08 (2H, s), 4.20 (2H, s), 3.58 (2H, t, J=5.85 Hz), 3.16 (6H, s), 2.42 (2H, t, J=6.22 Hz), 1.95 (2H, t, J=6.59).
2-(2,2-Dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-(4-fluoro-2-methylsulfanyl-benzyl)-N-methyl-acetamide was reacted with paraformaldehyde and 2-morpholin-4-yl-ethylamine as described in Method 44B to give the title compound as a white solid (0.032 g, 76% yield). 1HNMR (300 MHz, CDCl3) δ: 7.07 (1H, dd, J=9.52, 6.23 Hz), 6.92 (1H, dd, J=9.52, 2.56 Hz), 6.82 (1H, td, J=8.06, 2.57 Hz), 4.61 (2H, s), 4.27 (2H, s), 3.95–3.90 (8H, m), 3.76–3.72 (2H, m), 3.39 (2H, t, J=5.85 Hz), 2.97 (3H, s), 2.47 (3H, s).
2-(2,2-Dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-(4-fluoro-2-methylsulfanyl-benzyl)-N-methyl-acetamide was reacted with paraformaldehyde and amino-ethanol as described in Method 44 B to give the title compound as a white solid (0.28 g, 79% yield). 1HNMR (300 MHz, MeOD) δ: 7.21 (1H, dd, J=8.42, 5.86 Hz), 7.06 (1H, dd, J=9.88, 2.57 Hz), 6.88 (1H, td, J=8.41, 2.56 Hz), 4.69 (2H, s), 4.23 (2H, s), 3.72 (2H, t, J=10.24, 5.12H), 3.61–3.57 (2H, m), 3.02 (3H, s), 2.50 (3H, s).
2-(2,2-Dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-(4-fluoro-2-methanesulfinyl-benzyl)-N-methyl-acetamide was reacted with paraformaldehyde and 2-morpholin-4-yl-ethylamine as described in Method 44 B to give the title compound as a yellow solid (0.04 g, 88% yield). 1HNMR (300 MHz, MeOD) δ: 7.72 (1H, dd, J=8.42, 2.56 Hz), 7.49 (1H, dd, J=8.79, 5.13 Hz), 7.32–7.26 (1H, m), 4.19 (2H, s), 4.09–3.46 (10H, m), 3.30–3.21 (2H, m), 3.12 (3H, s), 2.81 (3H, s).
2-(2,2-Dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-(4-fluoro-2-methanesulfinyl-benzyl)-N-methyl-acetamide was reacted with paraformaldehyde and amino-ethanol as described in Method 44B to give the title compound as a colorless oil (0.015 g, 40% yield). 1HNMR (300 MHz, MeOD) δ: 7.71 (1H dd, J=8.42, 2.56 Hz), 7.47 (1H, dd, J=8.42, 5.12 Hz), 7.31 (1H, td, J=8.41, 2.56 Hz), 4.95 (2H, s), 4.23 (2H, s), 3.73 (2H, t, J=5.12 Hz), 3.59 (2H, t, J=4.76 Hz), 3.11 (3H, s), 2.80 (3H, s).
2-(2,2-Dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-(4-fluoro-2-methanesulfonyl-benzyl)-N-methyl-acetamide was reacted with paraformaldehyde and 2-morpholin-4-yl-ethylamine as described in Method 44B to give the title compound as a white solid (0.032 g, 69% yield). 1HNMR (300 MHz, DMSO) δ: 9.65 (1H, bs), 7.74 (1H, dd, J=8.41, 2.56 Hz), 7.64–7.59 (1H, m), 7.43–7.38 (1H, m), 4.99 (2H, s), 4.15 (2H, s), 4.00–3.53 (10H, m), 3.43 (2H, s), 3.38 (3H, s), 3.11 (3H, s).
2-(2,2-Dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-(4-fluoro-2-methanesulfonyl-benzyl)-N-methyl-acetamide was reacted with paraformaldehyde and amino-ethanol as described in Method 44 B to give the title compound as a colorless oil (0.02 g, 41% yield). 1HNMR (300 MHz, MeOD) δ: 7.77 (1H, dd, J=8.42, 2.57 Hz), 7.56–7.45 (2H, m), 5.10 (2H, s), 4.26 (2H, s), 3.73 (2H, bs), 3.59 (2H, bs), 3.23 (6H, s).
To methoxylamine hydrochloride (20 g, 0.24 mol) in water (200 mL) and THF (74 mL) was added sodium acetate (16.3 g, 0.2 mol) followed by 3,4-dichlorobenzaldehyde (25 g, 0.14 mol). The mixture was stirred at room temp for 6 h and diluted with diethyl ether. The aqueous phase was extracted with ethyl acetate, dried (sodium sulfate) and concentrated to give a colorless oil. The oil was dissolved in glacial acetic acid (200 mL) and cooled to 0° C. Sodium cyanoborohydride (18.8 g, 0.26 mol) was added over 30 min. The mixture was stirred at room temp for 4 days, cooled to 0° C. and made basic with 10 N NaOH. The mixture was extracted with EtOAc (3×'s) and the combined organic extracts were washed with water and brine and concentrated to give an oil that solidifies upon standing. This residue was stirred in diethyl ether and the resulting title compound was filtered as a white solid (9.61 g, 33% yield). 1HNMR (300 MHz, DMSO) δ: 10.12 (1H, bs), 7.74 (1H, s), 7.67 (1H, d, J=8.42 Hz), 7.44 (1H, d, J=8.05 Hz), 4.21 (2H, s), 3.59 (3H, s).
A suspension of (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetic acid (4.37 g, 25.4 mmol) in benzene (30 mL) was refluxed for 1 h with oxalyl chloride (15 mL). The resulting solution was cooled and concentrated. The residue was dissolved in dichloromethane (30 mL) and cooled to 0° C. and N-(3,4-dichloro-benzyl)-O-methyl-hydroxylamine (5.3 g, 22 mmol) in dichloromethane (30 mL) and pyridine (18 mL) was added dropwise. The resulting mixture was stirred at room temp for 18 h. The mixture was diluted with 1N HCl and extracted with EtOAc. The organic phase was washed with 1N HCl, dried (sodium sulfate) and concentrated to give the title compound as a yellow solid (7.59 g, 83% yield). 1HNMR (300 MHz, CDCl3) δ: 7.44 (1H, s), 7.39 (2H, d, J=8.05 Hz), 7.19 (2H, d, J=8.05 Hz), 6.37 (1H, s), 4.75 (2H, s), 3.70 (3H, s), 1.74 (6H, s).
A mixture of N-(3,4-dichloro-benzyl)-2-(2,2-dimethyl-5-oxo-[1,3]dioxolane-4-ylidene)-N-methoxy-acetamide (1.0 g, 2.8 mmol) and potassium carbonate (0.39 g, 2.8 mmol) in MeOH (20 mL) was stirred at room temp for 1 h. The suspension was diluted with EtOAc and washed with 1N HCl followed by brine. The organic phase was dried (sodium sulfate) and concentrated. The title compound was purified by flash chromatography eluting with 100% hexane followed by 25% EtOAc/hexane to give a white solid (0.3107 g, 33% yield). 1HNMR (300 MHz, CDCl3) δ: 13.28 (1H, s), 7.41 (2H, m), 7.15 (1H, dd, J=9.52, 1.47 Hz), 6.45 (1H, s), 4.77 (2H, s), 3.89 (3H, s), 3.72 (3H, s).
Amine (0.1 mmol) was reacted with paraformaldehyde according to the method described for the preparation of compound 86-A. This was combined with 3-[(3,4-dichloro-benzyl)-methoxy-carbamoyl]-2-hydroxy-acrylic acid methyl ester (0.036 g, 0.1 mmol), in MeOH (1.0 mL) and the resulting mixture stirred at 55° C. for 1 h. The resulting solution was cooled and purified by preparative HPLC (YMC Combiprep ODS-A, 30 mm×50 mm, MeOH/H2O/0.1% TFA) to give the title compounds. Compounds were evaluated by LC/MS (Waters XTERRA, 4.6 mm×30 mm, MeOH/H2O/0.1% TFA, 10%–90% MeOH, 2 min gradient, 5 mL/min flow rate; ESI+).
3-[(3,4-Dichloro-benzyl)-methoxy-carbamoyl]-2-hydroxy-acrylic acid methyl ester, paraformaldehyde and 4-amino-butanol were reacted according to the method described for compounds 792–808 to give the tile compound as a yellow oil (0.015 g, 36% yield). 1HNMR (300 MHz, CDCl3) δ: 7.43–7.40 (2H, m), 7.18 (1H, dd, J=8.42,1.83 Hz), 4.81 (2H, s), 4.14 (2H, s), 3.74 (3H, s), 3.63 (2H, t, J=6.40 Hz), 3.51 (2H, t, J=4.01 Hz), 1.68–1.53 (4H, M), 1.43–1.33 (2H, m).
To a solution of 4-amino butyric acid (0.093 g, 0.90 mmol) in acetic acid (0.5 mL) at 55° C. was added paraformaldehyde (0.027 g, 0.90 mmol). After stirring for 10 min, 3-[(3,4-dichloro-benzyl)-methoxy-carbamoyl]-2-hydroxy-acrylic acid methyl ester (0.2978 g, 0.89 mmol) was added and the mixture was stirred at 55° C. for 1 h. The clear yellow solution was cooled, concentrated and purified by preparative HPLC (C18, ODS-A, S-75 μm, 10%–40% acetonitrile/water/0.5% HCl) to give the title compound as a yellow foam (0.129 g, 34% yield). HRMS (M+H) calcd for C17H19N2Cl2O6: 417.06203; found: 417.0607.
The general method for the synthesis of compounds 811–814 is illustrated in Scheme XIV.
O-Methyl-N-naphthalen-1-ylmethyl-hydroxylamine was reacted with (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetic acid according to the method described for the preparation of compound 90-B to give the title compound as a pale yellow solid (0.826 g, 56% yield). 1HNMR (300 MHz, CDCl3) δ: 8.18 (1H, d, J=8.05 Hz), 7.85 (2H, t, J=9.15 Hz), 7.56–7.41 (4H, m), 6.40 (1H, s), 5.30 (2H, s), 3.43 (3H, s), 1.77 (6H, s).
Compound 93-B was prepared using the same procedure described for the preparation of compound 90-C and isolated as an orange powder (0.022 g, 45% yield). 1HNMR (300 MHz, CDCl3) δ: 8.09 (1H, d, J=8.05 Hz), 7.87 (2H, t, J=7.68 Hz), 7.57–7.41 (4H, m), 5.35 (2H, s), 4.16 (2H, s), 3.56 (3H, s), 3.54 (2H, t, J=6.33 Hz), 2.38 (2H, t, J=6.58 Hz), 1.91 (2H, t, J=6.22 Hz).
As illustrated in Scheme XIV, compounds 811–814 were prepared from 2-hydroxy-3-(methoxy-naphthalen-1-ylmethyl-carbamoyl)-acrylic acid methyl ester, paraformaldehyde and an amine, I-7 using the same method described for the preparation of compounds 792–808.
2-Hydroxy-3-(methoxy-naphthalen-1-ylmethyl-carbamoyl)-acrylic acid methyl ester, paraformaldehyde and 4-amino-butyric acid were reacted using the method described for the synthesis of Compound 810 to give the title compound as an orange powder (0.0217 g, 45% yield). 1HNMR (300 MHz, CDCl3) δ: 8.09 (1H, d, J=8.05 Hz), 7.87 (2H, t, J=768 Hz), 7.57–7.41 (4H, m), 5.35 (2H, s), 4.16 (2H, s), 3.56 (3H, s), 3.54 (2H, t, J=6.33 Hz), 2.38 (2H, t, J=6.58 Hz) 1.91 (2H, t, J=6.22 Hz).
A 250 mL round bottom flask was charged with 4-fluoro-2-methylsulfanylbenzaldehyde (5.106 g, 30 mmol), O-methylhydroxylamine hydrochloride (3.758 g, 45 mmol), sodium acetate (6.124 g, 45 mmol), water (50 mL) and THF (75 mL). The reaction mixture was stirred at room temperature for 4 h and taken up into ether (300 mL), washed with water (2×50 mL) and brine (50 mL). The organic phase was dried over anhydrous Na2SO4, filtered and concentrated to give 4-fluoro-2-methylsulfanyl-benzaldehyde-O-methyl-oxime (5.892 g, 99%) as a white solid.
To a stirred solution of 4-fluoro-2-methylsulfanyl-benzaldehyde-O-methyl-oxime (5.982 g, 29.58 mmol) in CH2Cl2 (30 mL) was added trifluoroacetic acid (30 mL) followed by triethylsilane (14 mL, 90 mmol). After stirring for 6 h, the reaction mixture was concentrated and the resulting residue was taken up in saturated aqueous NaHCO3 (100 mL) and extracted with ethyl acetate (3×75 mL). The combined organic extracts were dried over anhydrous Na2SO4, filtered, and concentrated to give a viscous oil. The crude product was re-dissolved in anhydrous ether and 2M HCl/ether w (18 mL) was added. The resulting white solid was filtered and dried to yield the desired product as the HCl salt (6.666 g, 93% yield). 1HNMR (500 MHz, DMSO-d6) δ: 11.44 (1H, br s), 7.66–7.59 (1H, m), 7.24 (1H, dd, J=10.1, 2.1 Hz), 7.08 (1H, td, J=8.6, 2.4 Hz), 4.85 (2H, s), 3.82 (3H, s), 2.53 (3H, s). MS calcd for C9H13FNOS (M+H): 202.07. found: 202.07.
To a stirred solution of N-(4-fluoro-2-methylsulfanyl-benzyl)-O-methyl-hydroxylamine hydrochloride (1.19 g, 5 mmol) and (2,2-Dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl chloride (0.953 g, 5 mmol) in CH2Cl2 (20 mL) was added diisopropyethylamine (1.9 mL, 11 mmol) at room temperature. After 1 h, the reaction mixture was concentrated and the resulting residue was taken up into ether (150 mL), washed with 1N HCl (10 mL), water (20 mL) and brine (20 mL). The organic layer was dried over anhydrous Na2SO4, filtered and concentrated to give product as a white solid (1.77 g, 99%). 1HNMR (300 MHz, CDCl3) δ: 7.28–7.23 (1H, m), 6.90 (1H, dd, J=9.5, 2.6 Hz), 6.78 (1H, td, J=8.4, 2.6 Hz), 6.37 (1H, s), 4.88 (2H, s), 3.62 (3H, s), 2.44 (3H, s), 1.71 (6H, s). MS calcd for C16H18FNO5S Na (M+Na): 378.078; found: 378.18.
2(2,2-Dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-(4-fluoro-2-methylsulfanyl-benzyl)-N-methoxy-acetamide was reacted with methanol to give 3-[(4-fluoro-2-methylsulfanyl-benzyl)-methoxy-carbamoyl]-2-hydroxy-acrylic acid methyl ester as a white sticky solid (0.0807 g, 88% yield). This was then reacted with paraformaldehyde and 4-amino butyric acid following the procedure described for Compound 810 to give the title compound as an orange solid (0.0360 g, 64% yield). 1HNMR (300 MHz, DMSO) δ: 12.10 (1H, bs), 11.35 (1H, bs), 7.30 (1H, m), 7.14 (1H, dd, J=12.44, 2.20 Hz), 6.97 (1H, td, J=8.41, 2.65 Hz), 4.86 (2H, s), 4.18 (2H, s), 3.69 (3H, s), 3.42 (2H, t, J=6.73 Hz), 2.50 (3H, t, J=1.83 Hz), 2.22 (2H, t, J=6.73 Hz), 1.78 (2H, m).
MCPBA (0.49 g, 1.41 mmol) was added to a solution of 2-(2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-(4-fluoro-2-methylsulfanyl-benzyl)-N-methoxy acetamide (0.50 g, 1.41 mmol) in dichloromethane (10 mL) and the mixture was stirred at room temp for 4 h. After diluting with EtOAc and washing with saturated NaHSO4, NaHCO3 and brine, the organic phase was dried (sodium sulfate) and concentrated. The title compound was purified by flash chromatography eluting with 100% hexane to 100% EtOAc to give a white foamy solid (0.4789 g, 92% yield). 1HNMR (300 MHz, CDCl3) δ: 7.75 (1H, dd, J=8.42, 2.92 Hz), 7.49 (1H, dd, J=8.42, 5.12 Hz), 7.15 (1H, td, J=8.42, 2.93 Hz), 6.34 (1H, s), 4.88 (2H, d, J=8.05 Hz), 3.70 (3H, s), 2.76 (3H, s), 1.74 (6H, s).
2-(2,2-Dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-(4-fluoro-2-methanesulfinyl-benzyl)-N-methoxy-acetamide was reacted with methanol using the procedure described for the synthesis of compound 90-C to yield 3-[(4-fluoro-2-methanesulfinyl-benzyl)-methoxy-carbamoyl]-2-hydroxy-acrylic acid methyl ester as a white sticky solid (0.1020 g, 100% yield). This crude was reacted with paraformaldehyde and 4-amino-butyric acid using the procedure described for Compound 810 to give the title compound as an orange solid (0.0360 g, 64% yield). 1HNMR (300 MHz, CDCl3) δ: 7.75 (1H, dd, J=8.05, 2.56 Hz), 7.48 (1H, dd, J=8.42, 4.76 Hz), 7.21 (1H, td, J=8.05, 2.56 Hz), 4.96 (2H, q), 4.19 (2H, s), 3.78 (3H, s), 3.58 (2H, td, J=6.95, 1.83 Hz), 2.82 (3H, s), 2.39 (2H, t, J=6.95 Hz), 1.97–1.92 (2H, m).
2-(2,2-Dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-(4-fluoro-2-methylsulfanyl-benzyl)-N-methoxy-acetamide was reacted with 2 equivalents of MCPBA using the method described for the synthesis of Compound 96-A to give the title compound as a white solid (0.263 g, 48% yield). 1HNMR (300 MHz, CDCl3) δ: 7.74 (1H, dd, J=8.06, 2.57 Hz), 7.60 (1H, dd, J=8.78, 5.12 Hz), 7.27 (1H, td, 7.68, 2.56 Hz), 6.45 (1H, s), 5.26 (2H, s), 3.81 (3H, s), 3.22 (3H, s), 1.74 (6H, s).
2-(2,2-Dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-(4-fluoro-2-methanesulfonyl-benzyl)-N-methoxy-acetamide was reacted with methanol using the procedure described for the synthesis of compound 90-C to yield 3-[(4-fluoro-benzyl)-methoxy-carbamoyl]-2-hydroxy-acrylic acid methyl ester as a white sticky solid (0.102 g, 100% yield). This was reacted with paraformaldehyde and 4-amino-butyric acid using the procedure described for Compound 810 to give the title compound as an orange solid (0.018 g, 14% yield). 1HNMR (300 MHz, CDCl3) δ: 7.78 (1H, dd, J=8.05, 2.56 Hz), 7.59–7.54 (1H, m), 7.36–7.29 (1H, m), 5.35 (2H, s), 4.25 (2H, s), 3.83 (3H, s), 3.60 (2H, t, J=6.95 Hz), 3.22 (3H, s), 2.43 (2H, t, J=6.95 Hz), 2.00–1.96 (2H, m).
2-(2,2-Dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-(4-fluoro-2-methylsulfanyl-benzyl)-N-methoxy-acetamide was reacted with paraformaldehyde and 2-morpholin-4-yl-ethylamine using the method described for the preparation of Compound 767 to give the title compound as a white solid (0.012 g, 18% yield). 1HNMR (300 MHz, CDCl3) δ: 7.27–7.22 (1H, m), 6.94 (1H, dd, J=9.51, 2.56 Hz), 6.83 (1H, td, J=8.42, 2.56 Hz), 4.95 (2H, s), 4.28 (2H, s), 3.97–3.75 (8H, m), 3.68 (3H, s), 3.40 (2H, t, J=5.85 Hz), 2.95 (2H, bs), 2.49 (3H, s).
2-(2,2-Dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-(4-fluoro-2-methylsulfanyl-benzyl)-N-methoxy-acetamide was reacted with paraformaldehyde and amino-ethanol using the method described for the synthesis of Compound 767 to give the title compound as a yellow oil (0.021 g, 56% yield). 1HNMR (300 MHz, MeOD) δ: 7.35–7.29 (1H, m), 7.08 (1H, d, J=8.05 Hz), 6.92–6.84 (1H, m), 4.99 (2H, s), 4.39 (2H, s), 3.76–3.70 (5H, m), 3.63–360 (2H, m), 2.51 (3H, s).
2-(2,2-Dimethyl-5-oxo-[1,2]dioxolan-4-ylidene)-N-(4-fluoro-2-methanesulfinyl-benzyl)-N-methoxy-acetamide was reacted with paraformaldehyde and 2-morpholin-4-yl-ethylamine using the method described for the preparation of Compound 767 to give the title compound as a yellow solid (0.045 g, 99% yield). 1HNMR (300 MHz, MeOD) δ: 7.27 (1H, dd, J=8.42, 2.56 Hz), 7.62 (1H, dd, J=8.79, 5.49 Hz), 7.32 (1H, td, J=8.41, 2.93 Hz), 5.06 (2H, s), 4.03–3.92 (4H, m), 3.83 (3H, s), 3.80–3.63 (4H, m), 3.51–3.46 (2H, m), 3.19 (2H, bs), 2.84 (3H, s).
2-(2,2-Dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-(4-fluoro-2-methanesulfonyl-benzyl)-N-methoxy-acetamide was reacted with paraformaldehyde and amino-ethanol using the method described for the preparation of compound 767 to give the title compound as a colorless oil (0.015 g, 40% yield). 1HNMR (300 MHz, MeOD) δ: 7.71 (1H, dd, J=8.78, 2.93 Hz), 7.61 (1H, dd, J=8.78, 5.12 Hz), 7.31 (1H, td, J=8.42, 2.56 Hz), 5.05 (2H, d, J=9.15 Hz), 4.38 (2H, s), 3.81 (3H, s), 3.74 (2H, t, J=5.12 Hz), 3.60 (2H, t, J=5.49 Hz), 2.84 (3H, s).
2-(2,2-Dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-(4-fluoro-2-methanesulfonyl-benzyl)-N-methoxy-acetamide was reacted with paraformaldehyde and 2-morpholin-4-yl-ethylamine using the method described for the preparation of compound 767 to give the title compound as a white solid (0.021 g, 45% yield). 1HNMR (300 MHz, MeOD) δ: 7.75 (1H, dd, J=8.78, 2.93 Hz), 7.63–7.54 (2H, m), 5.34 (2H, s), 4.29 (2H, s), 4.01–3.99 (2H, m), 3.84–3.81 (2H, m), 3.77 (3H, s), 3.63–3.45 (6H, m), 3.39 (3H, s), 3.16–3.08 (2H, m).
2-(2,2-Dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-(4-fluoro-2-methanesulfonyl-benzyl)-N-methoxy-acetamide was reacted with paraformaldehyde and amino-ethanol using the method described for the preparation of compound 767 to give the title compound as a colorless oil (0.100 g, 25% yield). 1HNMR (300 MHz, MeOD) δ: 7.77 (1H, dd, J=8.42, 2.56 Hz), 7.66 (1H, dd, J=8.79, 5.13 Hz), 7.45 (1H td, J=8.42, 2.93 Hz), 5.41 (2H, s), 4.46 (2H, s), 3.84 (3H, s), 3.76 (2H, t, J=5.12 Hz), 3.63 (2H, t, J=5.13 Hz), 3.28 (3H, s).
To a stirred suspension of 3,4-dichlorobenzylamine (17.6 mg, 0.1 mmol) and resin bound morpholine (100 mg, 2.5–4.0 mmol/1 g) in CH2Cl2 (2 mL) was added (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl chloride (20 mg, 0.11 mmol) and the mixture stirred for 1 h. The mixture filtered and the filtrate concentrated to give the product as a white solid (33 mg, 100% yield). 1H NMR (500 MHz, CDCl3) δ: 7.39 (2H, m), 7.15 (1H, m), 6.60 (1H, s), 5.89 (1H, s), 4.50 (2H, d, J=6.0 Hz), 1.74 (6H, s). MS calcd for C14H14Cl2NO4 [M+H]+: 330.03. found: 330.1.
2-Aminoethanol (0.8712 g, 10.0 mmol) and paraformaldehyde (0.3 g, 10.0 mmol) were stirred in methanol (50 mL) at 55° C. until the solids dissolved (approximately 20 min). The solution was cooled and used without further purification
A mixture of N-(3,4-dichloro-benzyl)-2-(2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetamide (66 mg, 0.2 mmol) and a 0.2 mmol solution of methylene-(2-morpholin-4-yl-ethyl)-amine in methanol (1.5 mL, 0.3 mmol) was heated at 70° C. for 1.5 h. The reaction mixture was then cooled to room temperature and purified by preparative HPLC using methanol/water (0.1% TFA) as eluent. The fractions containing the product were combined and concentrated to give a white powder (23.3 mg, 22% yield). 1H NMR (500 MHz, DMSO) δ: 9.50 (1H, br s), 8.08 (1H, t, J=6.2 Hz), 7.59 (1H, d, J=8.2 Hz), 7.53 (1H, d, J=1.8 Hz), 7.28 (1H, dd, J=8.2, 1.8 Hz), 4.42 (2H, d, J=6.1 Hz), 4.04 (2H, s), 4.02–3.95 (2H, m), 3.81–3.77 (2H, m), 3.73–3.37 (6H, m), 3.15–3.06 (2H, m). HRMS calcd for C18H22Cl2N3O4 (M+H): 414.09875. found: 414.0987.
To a stirred suspension of 3,4-difluorobenzylamine (0.43 mg, 0.3 mmol) and resin bound morpholine (200 mg, 2.5–4.0 mmol/1 g) was added a solution of (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl chloride (60 mg, 0.31 mmol) in CH2Cl2 (3 mL) and the mixture stirred for 1 h. The mixture was filtered and the filtrate concentrated to give the product as a white solid (85 mg, 95% yield). MS calcd for C14H13F2NO4Na (M+Na): 320.29; found: 320.27.
A mixture of N-(3,5-difluoro-benzyl)-2-(2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetamide (60 mg, 0.2 mmol) and a 0.2 mmol solution of methylene-(2-morpholin-4-yl-ethyl)-amine in methanol (1.25 mL, 0.25 mmol) was heated at 70° C. for 1 h. The reaction mixture was cooled to room temperature and purified by preparative HPLC using methanol/water (0.1% TFA) as the eluent. The fractions containing the product were combined and concentrated to give the title compound as a white powder (22 mg, 22% yield). 1H NMR (500 MHz, DMSO) δ: 11.95 (1H, br s), 9.54 (1H, br s), 8.08 (1H, t, J=6.1 Hz), 7.12–7.08 (1H, m), 7.00–6.94 (2H, m), 4.45 (2H, d, J=6.1 Hz), 4.05 (2H, s), 4.02–3.94 (2H, m), 3.83–3.77 (2H, m), 3.69–3.38 (6H, m), 3.16–3.05 (2H, m). HRMS calcd for C18H20F2N3O4 (M−H): 380.1422. found: 380.1422.
To a stirred suspension of 4-fluorobenzylamine (37.5 mg, 0.3 mmol) and resin bound morpholine (200 mg, 2.5–4.0 mmol/1 g) was added a solution of (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl chloride (60 mg, 0.31 mmol) in CH2Cl2 (3 mL) and the mixture shaken for 1 h. The mixture was filtered and the filtrate concentrated to give the title product as a white solid (84.4 mg, 100% yield). MS calc for C14H14FNO4 (M+Na): 302.29; found: 302.29.
A mixture of 2-(2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-(4-fluoro-benzyl)-acetamide (77 mg, 0.275 mmol) and a 0.2 mmol solution of methylene-(2-morpholin-4-yl-ethyl)-amine in methanol (1.5 mL, 0. mmol) was heated at 70° C. for 1 h. The reaction mixture was cooled to room temperature and purified by preparative HPLC using methanol/water (0.1% TFA) as the eluent. The fractions containing the product were combined and concentrated to give the title compound as a pale yellow powder (42 mg, 32% yield). 1H NMR (500 MHz, DMSO) δ: 11.92 (1H, br s), 9.48 (1H, br s), 7.95 (1H, t, J=4.9 Hz), 7.34–7.31 (2H, m), 7.17–7.13 (2H, m), 4.42 (2H, d, J=4.9 Hz), 4.04 (2H, s), 4.01–3.95 (2H, m), 3.80–3.77 (2H, m), 3.66–3.39 (6H, m), 3.15–3.06 (2H, m). HRMS calcd for C18H21FN3O4 (M+H): 362.1516. found: 362.1505.
To a stirred suspension of 3,4-difluorobenzylamine (0.43 mg, 0.3 mmol) and resin bound morpholine (200 mg, 2.5–4.0 mmol/1 g) was added a solution of (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl chloride (60 mg, 0.31 mmol) in CH2Cl2 (3 mL) and the mixture stirred for 1 h. The mixture was filtered and the filtrate concentrated to give the title product as a white solid (81.8 mg, 92% yield). MS calcd for C14H13F2NO4Na (M+Na): 320.29. found: 320.27.
A mixture of N-(3,4-difluoro-benzyl)-2-(2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetamide (58 mg, 0.195 mmol) and a 0.2 mmol solution of methylene-(2-morpholin-4-yl-ethyl)-amine in methanol (1.1 mL, 0.22 mmol) was heated at 70° C. for 1 h. The reaction mixture was cooled to room temperature and purified by preparative HPLC using methanol/water (0.1% TFA) as the eluent. The fractions containing the product were combined and concentrated to give the title compound as a hydroscopic yellow powder (30 mg, 31% yield). 1H NMR (500 MHz, DMSO) δ: 11.95 (1H, br s), 9.56 (1H, br s), 8.03 (1H, t, J=6.1 Hz), 7.41–7.30 (2H, m), 7.15–7.12 (1H, m), 4.41 (2H, d, J=6.1 Hz), 4.04 (2H, s), 4.02–3.93 (2H, m), 3.82–3.77 (2H, m), 3.65–3.51 (4H, m), 3.45–3.41 (2H, m), 3.15–3.06 (2H, m). HRMS calcd for C18H20F2N3O4 (M+H): 380.1422. found: 380.1424.
To a stirred solution of 3,4-dichlorobenzaldehyde (3.50 g, 20 mmol) and isopropylamine (2.40 g, 40 mmol) in methanol (20 mL) was added a freshly prepared solution zinc chloride (1.36 g, 10 mmol) and sodium cyanoborohydride (1.25 g, 20 mmol) in methanol (50 mL) at room temperature. The resulting clear reaction mixture was stirred overnight, then concentrated. The residue was taken up into aq NaOH (0.2N, 100 mL), extracted with ethyl acetate (3×100 mL). The combined organic extracts were dried over Na2SO4, filtered and concentrated to give viscous yellow oil. The residue was dissolved into ether and converted to hydrochloride salt (4.75 g, 93% yield). 1H NMR (500 MHz, DMSO) δ: 9.54 (2H, s), 8.00 (1H, d, J=2 Hz), 7.70 (1H, d, J=8 Hz), 7.65 (1H, m), 4.13 (2H, t, J=6 Hz), 3.25–3.22 (1H, m), 1.30 (3H, d, J=6.5 Hz). MS calcd for C10H14Cl2N (M+H): 218.04. found: 218.04.
To a stirred mixture of (3,4-dichloro-benzyl)-isopropyl-amine; compound hydrochloride (260 mg, 1.024 mmol) and (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl chloride (190 mg, 1.0 mmol) in CH2Cl2 (20 mL) at room temperature was added diisopropylethylamine (350 μL, 2.0 mmol). After 30 min, the reaction mixture was diluted with ethyl acetate (50 mL), washed with dilute aqueous HCl (1×5 mL), water (1×10 mL), and saturated aq. NaCl (1×5 mL). The organic layer was dried over Na2SO4, filtered and concentrated to give viscous yellow oil. Purification on a silica gel column using hexanes/ethyl acetate (3:2) gave pure product as a viscous yellow oil (372 mg, 93% yield). 1H NMR (500 MHz, CDCl3) δ: 7.41–7.30 (2H, m), 7.11 (0.67H, d, J=10.1 Hz), 7.10 (0.33H, d, J=10.1 Hz), 6.22 (0.67H, s), 5.87 (0.33H, s), 4.87–4.81 (0.33H, m), 4.51 (1.34H, s), 4.47 (0.66H, s), 4.30–4.24 (0.67H, m), 1.73 (4H, s), 1.71 (2H, s), 1.21 (4H, d, J=6.2 Hz), 1.11 (2H, d, J=6.7 Hz). MS calcd for C17H20Cl2NO4 (M+H): 372.24. found: 372.18.
Compound 829 was prepared according to the methods described in the previous examples to yield 217 mg (47% yield) of the title compound as the corresponding TFA salt. 1H NMR (300 MHz, DMSO) δ: 11.02 (1H, s), 9.62 (1H, br s), 7.57 (1H, d, J=8.1 Hz), 7.48 (1H, s), 7.22 (1H, d, J=7.7 Hz), 4.55 (2H, s), 4.33–4.24 (1H, br m), 4.11 (2H, s), 4.02–3.96 (2H, br m), 3.77 (2H, s), 3.64–3.54 (4H, m), 3.42 (2H, br s), 3.19–3.04 (2H, br m), 1.12 (6H, d, J=6.6 Hz). HRMS calcd for C21H28Cl2N3O4 (M+H): 456.1457. found: 456.1470.
To a stirred solution of 3,4-dichlorobenzaldehyde (3.50 g, 20 mmol) and 3-phenylpropylamine (5.40 g, 40 mmol) in methanol (20 mL) was added a freshly prepared solution of zinc chloride (1.36 g, 10 mmol) and sodium cyanoborohydride (1.25 g, 20 mmol) in methanol (50 mL) at room temperature. The resulting clear reaction mixture was stirred overnight, then concentrated. The residue was taken up into aq NaOH (0.2N, 100 mL) and extracted with ethyl acetate (3×100 mL). The combined organic extracts were dried over Na2SO4, filtered and concentrated to give a viscous yellow oil. The residue was purified on silica gel column using 30–40% ethyl acetate in hexanes. The product was dissolved in Et2O and treated with HCl (in ether) to form the corresponding hydrochloride salt as a precipitate which was isolated by filtration (5.40 g, 98% yield). 1H NMR (500 MHz, DMSO) δ: 9.66 (2H,s), 7.94 (1H, d, J=2.0 Hz), 7.69 (1H, d, J=8.5 Hz), 7.59 (1H, m), 7.30–7.18 (5H, m), 4.13 (2H, s), 2.89–2.5 (2H, br s), 2.65 (2H, t, J=7.5 Hz), 1.99 (2H, m). MS calcd for C16H17C2N (M+1): 294.08. found: 294.08.
To a stirred mixture of (3,4-dichloro-benzyl)-(3-phenyl-propyl)-amine; hydrochloride (330 mg, 1.0 mmol) and (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl chloride (206 mg, 1.08 mmol) in CH2Cl2 (10 mL) at room temperature was added diisopropylethylamine (386 μL, 2.2 mmol). After 30 min, the reaction mixture was diluted with ether (50 mL), washed with dilute aqueous HCl (1×5 mL), water (1×10 mL), and saturated aq. NaCl (1×5 mL). The organic layer was dried over Na2SO4, filtered and concentrated to give the desired amide (447 mg, 100% yield) as a viscous oil. 1H NMR (300 MHz, CDCl3) δ: 7.41–6.94 (8H, m), 6.08 (0.66H, s), 6.01 (0.34H, s), 4.53 (1.33H, s), 4.48 (0.67H, s), 3.41 (0.67H, t, J=7.5 Hz), 3.26 (1.33H, t, J=7.8 Hz), 2.61 (2H, t, J=7.3 Hz), 1.95–1.85 (2H, m), 1.71 (6H,s). MS calcd for C23H23Cl2NO4Na (M+Na): 470.09. Found: 470.09.
Compound 830 was prepared according to the methods described in the previous examples to yield 207 mg (38% yield) of the title compound as the corresponding TFA salt. 1HNMR (500 MHz, DMSO-d6) δ: 11.04 (1H, s), 9.64 (1H, s), 7.59 (1H, d, J=8.2 Hz), 7.50 (1H, s), 7.28–7.21 (3H, m), 7.17–7.11 (3H, m), 4.63 (2H, s), 4.07 (2H, s), 4.04–3.96 (2H, br s), 3.79–3.73 (2H, br s), 3.68–3.51 (4H, br m), 3.46–3.35 (4H, br m), 3.28 (1H, br s), 3.16–3.05 (2H, br s), 1.88–1.71 (2H, br s). HRMS calcd for C27H32Cl2N3O4 (M+H): 532.1770. found: 532.1758.
To a stirred solution of 3,4-dichlorobenzaldehyde (3.50 g, 20 mmol) and 3-methylbutylamine (2.62 g, 40 mmol) in methanol (20 mL) was added a freshly prepared solution zinc chloride (1.36 g, 10 mmol) and sodium cyanoborohydride (1.25 g, 20 mmol) in methanol (50 mL) at room temperature. The resulting clear reaction mixture was stirred overnight, then concentrated. The residue was taken up into aq NaOH (0.2N, 100 mL), extracted with ethyl acetate (3×100 mL). The combined organic extracts were dried over Na2SO4, filtered and concentrated to give viscous yellow oil. The residue was purified on silica gel column using 1:4, 3:7 and 2:3 ethyl acetate/hexanes as eluent. The product was dissolved in Et2O and treated with HCl (in ether) to form the corresponding hydrochloride salt as a precipitate which was isolated by filtration (4.924 g, 73% yield). 1H NMR (500 MHz, DMSO) δ: 9.57 (2H,s), 7.95 (1H, d, J=2 Hz), 7.70 (1H, d, J=8.5 Hz), 7.60 (1H, m), 4.13 (2H, s), 2.87–2.85 (2H, br m), 1.63–1.54 (3H, m), 0.86 (6H, d, J=6.5 Hz). MS calcd for C12H18Cl2N (M+H): 246.08. found: 246.31.
To a stirred mixture of (3,4-dichloro-benzyl)-(3-methyl-butyl)-amine hydrochloride (283 mg, 1.0 mmol) and (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl chloride (206 mg, 1.08 mmol) in CH2Cl2 (10 mL) at room temperature was added diisopropylethylamine (386 μL, 2.2 mmol). After 30 min, the reaction mixture was diluted with ether (50 mL), washed with dilute aqueous HCl (1×5 mL), water (1×10 mL), and saturated aq. NaCl (1×5 mL). The organic layer was dried over Na2SO4, filtered and concentrated to give viscous yellow oil. Purification on a Silica gel column using hexanes/ethyl acetate (20–40% EtOAc) as the eluent gave the title compound as a viscous yellow oil (400 mg, 100% yield). 1H NMR (500 MHz, CDCl3) δ: 7.42–7.26 (2H, m), 7.11 (0.67H, d, J=8.2 Hz), 7.01 (0.33H, d, J=8.2 Hz), 6.14 (0.67H, s), 6.00 (0.33H, s), 4.56 (1.34H, s), 4.52 (0.66H, s), 3.39–3.36 (0.67H, m), 3.29–3.26 (1.33H, m), 1.74 (4H, s), 1.70 (2H, s), 1.59–1.50 (1H, m), 1.47–1.42 (2H, m), 0.90–0.88 (6H, m). MS calcd for C19H24Cl2NO4 (M+H): 400.1. found: 400.07.
Compound 831 was prepared according to the methods described in the previous examples to yield 213 mg (38% yield) of the title compound as the corresponding TFA salt. 1HNMR (500 MHz, DMSO-d6) δ: 11.08 (1H, s), 9.73 (1H, s), 7.61 (1H, d, J=8.2 Hz), 7.52 (1H, s), 7.25 (1H, br s), 4.61 (2H, s), 4.11 (2H, s), 4.05–3.95 (2H, br m), 3.78 (2H, br s), 3.70–3.48 (4H, m), 3.46–3.31 (4H, br m), 3.26 (1H, br s), 3.17–3.02 (2H, br m), 1.57–1.27 (3H, br m), 0.80 (6H, br s). HRMS calcd for C23H32Cl2N3O4 (M+H): 484.177. found: 484.1793.
A mixture of paraformaldehyde (18 mg, 0.563 mmol) and 4-(2-aminoethyl)tetrahydropyran (65 mg, 0.5 mmol) in anhydrous methanol (3 mL) was warmed to 60° C. To the resulting clear homogeneous solution was added N-(3,4-dichloro-benzyl)-2-(2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-methyl-acetamide (172 mg, 0.5 mmol) and stirring continued for 18 h at 60° C. and an additional 6 h at room temperature. The crude reaction mixture was purified by preparative HPLC on a C18 column using water/methanol (0.1% TFA) as the eluent. The fractions containing the desired product were combined, the methanol removed under vacuum and the remaining aqueous solution extracted with ethyl acetate. The combined ethyl acetate extracts were dried over anhydrous Na2SO4, filtered and concentrated to give the title compound as a white solid (164 mg, 77% yield). 1HNMR (500 MHz, CDCl3) δ: 10.15 (1H, br s), 7.41 (1H, d, J=8.2 Hz), 7.35 (1H, s), 7.11 (1H, d, J=8.2 Hz), 4.60 (2H, s), 4.13 (2H, s), 3.93 (2H, dd, J=11, 3.7 Hz), 3.54 (2H, t, J=7.1 Hz), 3.34 (2H, t, J=11.9 Hz), 3.03 (3H, s), 1.65–1.24 (7H, m). HRMS calcd for C20H25Cl2N2O4 (M+H): 427.1191. found: 427.1179.
A mixture of paraformaldehyde (56 mg, 1.75 mmol) and 1-(2-aminoethyl)thiomorpholine (219 mg, 1.5 mmol) in anhydrous methanol (5 mL) was warmed to 60° C. To the resulting clear homogeneous solution was added N-(3,4-dichloro-benzyl)-2-(2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-methyl-acetamide (516 mg, 1.5 mmol) and stirring continued for 18 h at 60° C. and an additional 6 h at room temperature. The crude reaction mixture was purified on C18 column using water/acetonitrile (contains 0.05% TFA) as the eluent. The fractions containing the desired product were combined, concentrated and lyophilized to give the corresponding TFA salt of the title compound as a white solid (555 mg, 66% yield). 1HNMR (500 MHz, DMSO-d6) δ: 11.10 (1H, s), 9.53 (1H, br s), 7.62 (1H, d, J=8.2 Hz), 7.50 (1H, s), 7.28–7.20 (1H, br s), 4.59 (2H, s), 4.12 (2H, s), 3.85–3.73 (4H, m), 3.45–3.37 (2H, br s), 3.25–3.14 (2H, br m), 3.02–2.95 (2H, br m), 2.94–2.85 (4H, br m). HRMS calcd for C19H24Cl2N3O3S: 444.0915. found: 444.0909.
A mixture of paraformaldehyde (18 mg, 0.563 mmol) and 2-(2-aminoethylamino)ethanol (54 mg, 0.5 mmol) in anhydrous methanol (3 mL) was warmed to 60° C. To the resulting clear homogeneous solution was added N-(3,4-dichloro-benzyl)-2-(2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-methyl-acetamide (175 mg, 0.5 mmol) and the mixture stirred for 30 min at 60° C. The crude product was purified by preparative HPLC on a C18 column using water/methanol (0.1% TFA) as the eluent. The fractions containing the desired product were combined, concentrated and lyophilized to give the corresponding TFA salt of the title compound as a white powder (156 mg, 61%.yield). 1HNMR (500 MHz, CDCl3) δ: 9.08 (2H, br s), 7.39 (1H, d, J=8.2 Hz), 7.32 (1H, s), 7.08 (1H, d, J=8.2 Hz), 4.55 (2H, s), 4.24 (2H, s), 3.93–3.80 (5H, br m), 3.40 (2H, br s), 3.21 (2H, br s), 2.98 (3H, s). HRMS calcd for C17H22C2N3O4 (M+H): 402.0987. found: 402.1311.
To a stirred solution of 4-hydroxy-5-oxo-1-(2-thiomorpholin-4-yl-ethyl)-2,5-dihydro-1H-pyrrole-3-carboxylic acid (3,4-dichloro-benzyl)-methyl-amide (108 mg, 0.243 mmol) in water (20 mL) was added 30% hydrogen peroxide (0.3 mL). After stirring overnight the reaction mixture was lyophilized to give the title compound as a white powder. 1HNMR (500 MHz, CDCl3) δ: 7.42 (1H, d, J=8.5 Hz), 7.34 (1H, s), 7.10 (1H, d, J=8.2 Hz), 4.59 (2H, s), 4.28 (2H, s), 3.89 (2H, t, J=5.8 Hz), 3.74–3.70 (2H, m), 3.63 (2H, t, J=11.9 Hz), 3.29–3.24 (2H, m), 3.18 (2H, t, J=11.9 Hz), 3.07–2.99 (2H, m), 3.01 (3H, s). HRMS calcd for C19H24Cl2N3O4S (M+H): 460.0865. found: 460.0884.
To a stirred warm (approximately 60° C.) solution of 2-(2-aminoethylamino)ethanol (104 mg, 1.0 mmol) and paraformaldehyde (32 mg, 1.0 mmol) in anhydrous methanol was added N-(3,4-dichloro-benzyl)-2-(2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-methyl-acetamide (344 mg, 1 mmol). After 30 min, the mixture was cooled to room temperature and methyl bromoacetate (104 μL, 1.1 mmol) followed by K2CO3 was added. The resulting suspension was stirred overnight at 40° C., then filtered and purified by preparative HPLC on a C18 column using water/methanol (0.1% TFA) as the eluent. The fractions containing the desired product were combined, concentrated and lyophilized to yield the corresponding TFA salt of the title compound as a white powder (248 mg, 42% yield). 1HNMR (500 MHz, CDCl3) δ: 7.38 (1H, d, J=8.2 Hz), 7.32 (1H, s), 7.07 (1H, d, J=8 Hz), 4.56 (3H, s), 4.20 (2H, s), 4.01 (2H, s), 3.86–3.78 (4H, m), 3.59–2.55 (2H, m), 3.38–3.34 (2H, m), 3.01–2.94 (4H, m), 2.73 (3H, s). HRMS calcd for C20H26Cl2N3O5 (M+H): 474.1199. found: 474.1295.
To a stirred warm (60° C.) solution of ethyl 4-amino-1-piperidinecarboxylate (86.1 mg, 0.5 mmol) and paraformaldehyde (16 mg, 0.5 mmol) in methanol (2 mL) was added N-(3,4-dichloro-benzyl)-2-(2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-methyl-acetamide (175 mg, 0.5 mmol). After 2 h, the reaction mixture was cooled and purified using preparative HPLC on a C18 column using water/methanol-(0.1% TFA) as the eluent. The fractions containing the desired product were combined, concentrated and lyophilized to give the title compound as a white powder (64 mg, 27% yield). 1HNMR (500 MHz, CDCl3) δ: 10.16 (2H, br s), 7.40 (1H, d, J=8.2 Hz), 7.35 (1H, s), 7.10 (1H, d, J=8.0 Hz), 4.59 (2H, s), 4.34–4.18 (3H, m), 4.13 (2H, q, J=7.0 Hz), 4.09 (2H, s), 2.06 (2H, s), 2.87–2.80 (2H, br m), 1.82–1.76 (2H, br m), 1.63–1.55 (2H, m), 1.25 (3H, t, J=7.0 Hz). MS calcd for C21H26Cl2N3O5 (M+H): 470.36. found: 470.02.
To a stirred warm (60° C.) solution of 3-aminopiperidine dihydrochloride (150 mg, 0.867 mmol), paraformaldehyde (34 mg, 1.04 mmol) and triethylamine (0.3 mL, 2.14 mmol) in methanol (5 mL) was added N-(3,4-dichloro-benzyl)-2-(2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-methyl-acetamide (300 mg, 0.872 mmol). After 1 h, the reaction mixture was cooled and purified by preparative HPLC using a C18 column and water/methanol (0.001 mM HCl) as eluent. The fractions containing the desired product were combined, concentrated and lyophilized to give the corresponding hydrochloride salt of the title compound as a white powder (162 mg, 43% yield). 1HNMR (500 MHz, DMSO-d6) δ: 11.06 (1H, br s), 9.35 (1H, d, J=10.1 Hz), 9.10–8.99 (1H, m), 7.62 (1H, d, J=8.2 Hz), 7.52 (1H, s), 7.29–7.20 (1H, br s), 4.59 (2H, d, JAB=15.7 Hz), 4.34–3.88 (4H, m), 3.40–2.73 (6H, m), 1.90–1.74 (4H, m). HRMS calcd for C18H22Cl2N3O3 (M+H): 398.1038; found: 398.1042.
To a stirred warm (60° C.) solution of 2,2,2-trifluoroethylamine hydrochloride (100 mg, 1.0 mmol) paraformaldehyde (32 mg, 1.0 mmol) and triethylamine (0.14 mL, 1.0 mmol) in methanol (5 mL) was added N-(3,4-dichloro-benzyl)-2-(2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-methyl-acetamide (344 mg, 1.0 mmol). After 24 h, the reaction mixture was cooled and purified by preparative HPLC using a C18 column and water/methanol-(0.1% TFA) as eluent. The fractions containing the desired product were combined, concentrated and lyophilized to give the title compound as a white powder (81.4 mg, 21% yield). 1HNMR (500 MHz, CDCl3) δ: 10.05 (1H, br s), 7.42 (1H, d, J=8.2 Hz), 7.36 (1H, s), 7.11 (1H, d, J=8.2 Hz), 4.60 (2H, s), 4.32 (2H, s), 4.10 (2H, q, 8.9 Hz), 3.04 (3H, s). HRMS calcd for C15H12Cl2F3N2O3 (M−H): 395.0177. found: 395.0192.
To a stirred warm (60° C.) solution of 4-aminopiperidine (36 mg, 0.25 mmol) and paraformaldehyde (8 mg, 0.25 mmol) in methanol (1.5 mL) was added N-(3,4-dichloro-benzyl)-2-(2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-methyl-acetamide (93 mg, 0.27 mmol). After 1 h, the reaction mixture was cooled and purified by preparative HPLC on a C18 column using water/methanol-(0.1% TFA) as the eluent. The fractions containing the desired product were combined, concentrated and lyophilized to give the corresponding TFA salt of the title compound as white powder (46.3 mg, 36% yield). 1HNMR (300 MHz, CDCl3) δ: 9.65 (1H, br s), 9.17 (1H, br s), 7.42 (1H, d, J=8.1 Hz), 7.34 (1H, s), 7.10 (1H, d, J=8.1 Hz), 4.60 (2H, s), 4.40–4.29 (1H, m), 4.20 (2H, s), 3.65–3.52 (2H, m), 3.10–2.82 (2H, m), 3.04 (3H, s), 2.43–1.97 (4H, m). HRMS calcd for C18H22Cl2N3O3 (M+H): 398.1038. found: 398.1044.
The general method for the preparation of compounds 857–862 is illustrated in Scheme XVI.
To a stirred solution of 3,5-dichlorobenzaldehyde (9.0 g, 51 mmol) in methanol (100 mL) was added methanolic methylamine (2M, 100 mL, 200 mmol) and the resulting mixture stirred for 2 h at room temperature. To this was added a solution of ZnCl2 (3.402 g, 25 mmol) slowly and NaCNBH3 (3.142 g, 50 mmol) in methanol (100 mL). After 24 h, the reaction mixture was concentrated, and the resulting residue taken up in dilute aq NaOH (0.5 N, 200 mL) and extracted with CH2Cl2 (5×50 mL). The combined CH2Cl2 extracts were dried over anhydrous Na2SO4, filtered and concentrated to give a yellow viscous liquid. The crude product was dissolved in ether (200 mL) to which was added 29 mL of 2N HCl (in ether). The resulting white 3,5-dichlorobenzyl-methylamine hydrochloride salt was filtered and dried under vacuum to give 11.20 g (97% yield) of product. 1HNMR (300 MHz, DMSO-d6) δ: 9.48 (2H, s), 7.68 (3H, s), 4.13 (2H, s), 2.50 (3H, s). MS calcd for C8H10C12N (M+H): 190.02. found: 190.05.
To a stirred solution of (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl chloride (0.96 g, 5 mmol) in CH2Cl2 (20 mL) was added a solution of 3,5-dichlorobenzyl-methylamine hydrochloride (1.133 g, 5 mmol) and Et3N (2 mL) in CH2Cl2 (25 mL). The addition flask was rinsed with CH2Cl2 (5 mL) and added to the reaction mixture. After 2 h, the reaction mixture was concentrated and the resulting residue was triturated with ether (100 mL), filtered and concentrated to give the desired product as a viscous brown oil (1.70 g, 100% yield). 1HNMR (300 MHz, CDCl3) δ: 7.30–7.27 (1H, m), 7.16 (1.5H, br s), 7.06 (0.5H, s), 6.17 (0.66H, s), 6.07 (0.34H, s), 4.59 (1.33H, s), 4.53 (0.67H, s), 3.03 (2H, s), 2.97 (1H, s), 1.74 (4H, s), 1.70 (2H,s).
As illustrated in Scheme XVI, a 0.2 mM solution of amine, I-7, and paraformaldehyde in methanol (1 mL) are added to 0.1 mM solution of N-(3,5-dichloro-benzyl)-2-(2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-methyl-acetamide in methanol (2 mL). The mixture is warmed to 60° C. and kept at this temperature until the reaction is complete (1–24 h). The reaction mixture is purified by preparative HPLC on a C18 column using water/methanol-(0.1% TFA) as eluent. The fractions containing the desired product were combined, concentrated and lyophilized.
Reaction time 1 h. Obtained 47 mg (53% yield). 1HNMR (500 MHz, CDCl3) δ: 7.29 (1H, s), 7.13 (2H, s), 4.59 (2H, s), 4.16 (2H, s), 3.11 (3H, s), 3.03 (3H, s). HRMS calcd for C14H15Cl2N2O3 (M+H): 329.04598. found: 329.0456.
Reaction time 1 h. Obtained 39.8 mg (44% yield). 1HNMR (500 MHz, CDCl3) δ: 7.29 (1H, s), 7.14 (2H, s), 4.60 (2H, s), 4.17 (2H, s), 3.36 (2H, q, J=7.3 Hz), 3.04 (3H, s), 1.22 (3H, t, J=7.3 Hz). HRMS calcd for C15H17Cl2N2O3 (M+H): 343.0616. found: 343.0609.
Reaction time 24 h. Obtained 13.6 mg (14% yield). 1HNMR (500 MHz, CDCl3) δ: 7.29 (1H, s), 7.14 (2H, s), 4.65 (1H, t, J=4.6 Hz), 4.59 (2H, s), 4.56 (1H, t, J=4.6 Hz), 4.33 (2H, s), 3.84 (1H, t, J=4.6 Hz), 3.78 (1H, t, J=4.6 Hz), 3.04 (3H, s). HRMS calcd for C15H14Cl2FN2O3 (M−H): 359.0366. found: 359.0374.
Reaction time 1 h. Obtained 42.3 mg (41% yield) as the corresponding TFA salt. 1HNMR (500 MHz, CDCl3) δ: 9.66 (1H, s), 9.17 (1H, s), 7.30 (1H, s), 7.14 (2H, s), 4.60 (2H, s), 4.37 (1H, m), 4.21 (2H, s), 3.56–3.53 (2H, m), 3.08–3.03 (2H, m), 3.06 (3H, s), 2.27–2.19 (2H, m), 2.03–1.97 (2H, m). HRMS calcd for C18H22Cl2N3O3 (M+H): 398.1038. found: 398.1040.
Reaction time 1 h. Obtained 61.1 mg (56% yield) as the corresponding TFA salt. 1HNMR (500 MHz, CDCl3) δ: 9.15 (1H, br s), 7.29 (1H, s), 7.12 (2H, s), 4.57 (2H, s), 4.28 (2H, s), 4.06–3.72 (8H, m), 3.45–3.37 (2H, br s), 3.00 (3H, s), 2.98–2.91 (2H, m). HRMS calcd for C19H24Cl2N3O4 (M+H): 428.1144. found: 428.1135.
1HNMR (500 MHz, DMSO-d6) δ: 11.01 (1H, s), 7.53 (1H, s), 7.32 (2H, s), 4.60 (2H, s), 4.10 (2H, s), 3.64 (2H, t, J=5.35 Hz), 3.43 (3H, br s), 3.07–3.00 (4H, m). HRMS calcd for C15H18Cl2N3O3 (M+H): 358.0725. found: 358.0719.
To a stirred solution of 4-hydroxy-1-(2-methylamino-ethyl)-5-oxo-2,5-dihydro-1H-pyrrole-3-carboxylic acid (3,4-dichloro-benzyl)-methyl-amide (Compound 15) (150 mg, 0.367 mmol) in CH2Cl2 (5 mL) was added Et3N (0.14 mL, 1.0 mmol) followed by dimethylsulfamoyl chloride (53 μL, 0.5 mmol) at room temperature. After 4 h, the reaction mixture was concentrated and the resulting residue was dissolved in methanol and purified by preparative HPLC on a C18 column using water/methanol-(0.1% TFA) as eluent. The fractions containing the desired product were combined, concentrated and lyophilized to give the title compound as a white solid (134.3 mg, 62% yield). 1HNMR (500 MHz, CDCl3) δ: 10.38 (1H, br s), 7.42 (1H, d, J=8.2 Hz), 7.35 (1H, s), 7.10 (1H, d, J=8.2 Hz), 4.58 (2H, s), 4.29 (2H, s), 3.69 (2H, t, J=5.8 Hz), 3.43 (2H, t, J=5.8 Hz), 3.02 (3H, s), 2.81 (3H, s), 2.73 (6H, s). HRMS calcd for C18H25Cl2N4O5S (M+H): 479.0923. found: 479.0941.
To a stirred solution of 4-hydroxy-1-(2-methylamino-ethyl)-5-oxo-2,5-dihydro-1H-pyrrole-3-carboxylic acid (3,4-dichloro-benzyl)-methyl-amide (Compound 15) (150 mg, 0.367 mmol) in CH2Cl2 (5 mL) was added Et3N (0.14 mL, 1.0 mmol) followed by methanesulfonyl chloride (38 μL, 0.5 mmol) at room temperature. After 1 h, the reaction mixture was concentrated and the resulting residue was dissolved in methanol and purified by preparative HPLC on a C18 column using water/methanol-(0.1% TFA) as eluent. The fractions containing the desired product were combined, concentrated and lyophilized to give the title compound as a white powder (21.3 mg, 13% yield). 1HNMR (500 MHz, CDCl3) δ: 7.42 (1H, d, J=8.2 Hz), 7.35 (1H, s), 7.11 (1H, d, J=8.2 Hz), 4.58 (2H, s), 4.29 (2H, s), 3.71 (2H, t, J=5.8 Hz), 3.38 (2H, t, J=5.8 Hz), 3.02 (3H, s), 2.89 (3H, s), 2.78 (3H, s). HRMS calcd for C17H22Cl2N3O5S (M+H): 450.0657. found: 450.0658.
To a stirred solution of 4-hydroxy-1-(2-methylamino-ethyl)-5-oxo-2,5-dihydro-1H-pyrrole-3-carboxylic acid (3,4-dichloro-benzyl)-methyl-amide (Compound 15) (155 mg, 0.379 mmol) in CH2Cl2 (5 mL) was added Et3N (0.14 mL, 1.0 mmol) followed by methyl isocyanate (29 μL, 0.5 mmol) at room temperature. After 1 h, the reaction mixture was concentrated and the resulting residue was dissolved in methanol and purified by preparative HPLC on a C18 column using water/methanol (0.1% TFA) as eluent. The fractions containing the product were combined, concentrated and lyophilized to give the title compound as a white powder (107.6 mg, 52% yield). 1HNMR (500 MHz, CDCl3) δ: 11.56 (1H, br s), 7.41 (1H, d, J=8.2 Hz), 7.34 (1H, s), 7.10 (1H, d, J=8.2 Hz), 4.66 (1H, br s), 4.57 (2H, s), 4.28 (2H, s), 3.62 (2H, t, J=5.8 Hz), 3.56 (2H, t, J=5.8 Hz), 3.02 (3H, s), 2.89 (3H, s), 2.70 (3H, s). HRMS calcd for C18H23Cl2N4O4 (M+H): 429.1096. found: 429.1082.
To a stirred solution of 4-hydroxy-1-(2-methylamino-ethyl)-5-oxo-2,5-dihydro-1H-pyrrole-3-carboxylic acid (3,4-dichloro-benzyl)-methyl-amide (Compound 15) (153 mg, 0.374 mmol) in CH2Cl2 (5 mL) was added Et3N (0.14 mL, 1.0 mmol) followed by trimethylsilyl isocyanate (68 μL, 0.5 mmol) at room temperature. After 2 h, the reaction mixture was concentrated and the resulting residue was dissolved in methanol and purified by preparative HPLC on a C18 column using water/methanol-(0.1% TFA) as eluent. The fractions containing the product were combined, concentrated and lyophilized to yield the title compound as a white powder (120 mg, 61% yield). 1HNMR (500 MHz, CDCl3) δ: 10.37 (1H, br s), 7.41 (1H, d, J=8.2H), 7.34 (1H, s), 7.10 (1H, d, J=8.2 Hz), 5.78 (2H, br s), 4.56 (2H, s), 4.22 (2H, s), 3.64 (2H, t, J=5.2 Hz), 3.55 (2H, t, J=5.2 Hz), 2.99 (3H, s), 2.94 (3H, s). HRMS calcd for C17H21Cl2N4O4 (M+H): 415.094. found: 415.0941.
To a stirred solution of 4-hydroxy-1-(2-methylamino-ethyl)-5-oxo-2,5-dihydro-1H-pyrrole-3-carboxylic acid (3,4-dichloro-benzyl)-methyl-amide (Compound 15) (110 mg, 0.269 mmol) in CH2Cl2 (5 mL) was added Et3N (0.14 mL, 1.0 mmol) followed by a freshly prepared solution of t-butanol and chlorosulfonyl isocyanate (concentration=0.2 mM, 2 mL, 0.4 mmol) at room temperature. After 1 h, the reaction mixture was concentrated and the resulting residue dissolved in methanol and purified by preparative HPLC on a C18 column using water/methanol-(0.1% TFA) as eluent. The fractions containing the product were combined, concentrated and lyophilized to give the title compound as a white powder (77 mg, 43% yield). 1HNMR (500 MHz, CDCl3) δ: 7.68 (1H, s), 7.41 (1H d, J=8.2 Hz), 7.35 (1H, s), 7.10 (1H, d, J=8.2 Hz), 4.57 (2H, s), 4.26 (2H, s), 3.68 (2H, t, J=5.2 Hz), 3.54 (2H, t, J=5.2 Hz), 3.00 (3H, s), 2.92 (3H, s), 1.45 (9H, s). HRMS calcd for C21H27Cl2N4O7S (M+H): 549.0978. found: 549.0988.
Compound 852 was prepared according to the methods described in the previous examples. 1HNMR (500 MHz, DMSO-d6) δ: 10.98 (1H, br s), 7.62 (1H, d, J=8.24 Hz), 7.52 (1H, d, J=2.14 Hz), 7.25 (1H, d, J=7.33 Hz), 4.59 (2H, s), 4.10 (2H, s), 3.64 (5H, br s), 3.06–3.00 (4H, m). HRMS calcd for C15H18Cl2N3O3 (M+H): 358.0725. found: 358.0717.
Compound 853 was prepared according to the methods described in the previous examples. 1HNMR (500 MHz, CDCl3) δ: 11.96 (1H, br s), 8.92 (2H, s), 7.53 (1H, s), 7.31 (2H, s), 4.60 (2H, s), 4.11 (2H, s), 3.89 (3H, br s), 3.70 (2H, br s), 3.15 (2H, m), 2.55 (3H, t, J=5.19 Hz). HRMS calcd for C16H20Cl2N3O3 (M+H): 372.0882. found: 372.0884.
To a stirred solution of 4-hydroxy-1-(2-methylamino-ethyl)-5-oxo-2,5-dihydro-1H-pyrrole-3-carboxylic acid (3,5-dichloro-benzyl)-methyl-amide (82 mg, 0.2 mmol) and Et3N (0.084 mL, 0.6 mmol) in CH2Cl2 (3 mL) was added dimethylsulfamoyl chloride (0.028 mL, 0.26 mmol). After stirring for 2 h at room temperature, the reaction mixture was concentrated and purified by preparative HPLC using MeOH/Water as the eluent. The fractions containing the product were combined and concentrated to give the title compound as a paste (70 mg, 59% yield). 1HNMR (500 MHz, CDCl3) δ: 10.68 (1H, br s), 7.27 (1H, s), 7.13 (2H, s), 4.58 (2H, s), 4.29 (2H, s), 3.71–3.65 (2H, m), 3.45–3.40 (2H, m), 3.02 (3H, s), 2.80 (3H, s), 2.71 (6H, s). HRMS calcd for C18H25Cl2N4O5S (M+H): 479.0923. found: 479.0918.
Compound 855 was prepared according to the methods described in the previous examples. 1HNMR (500 MHz, CDCl3) δ: 7.84 (1H, d, J=8.24 Hz), 7.28 (1H, br s), 7.05–7.01 (1H, m), 4.52 (2H, s), 4.17 (2H, s), 4.03 (2H, s), 3.43 (3H, br s), 2.94 (3H, s). HRMS calcd for C15H14Cl2N3O4 (M−H): 370.0361; found: 370.0361.
Reaction of 2-(2,2-dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-methoxy-N-(4-methylbenzyl)-acetamide (0.30 g, 0.98 mmol) with the paraformaldehyde-N-(2-aminoethyl)morpholine adduct in methanol using a procedure similar to the one described in the preparation of compound 13 gave 0.17 g (45% yield) of the title compound as a solid. 1HNMR 400 MHz (CDCl3) δ (ppm); 2.35 (3H, s, CH3), 2.47 (4H, m, 2×NCH2), 2.56 (2H, t, J=6.2 Hz, NCH2), 3.62 (2H, t, J=6.2 Hz, NCH2), 3.66 (4H, m, 2×OCH2), 3.71 (3H, s, OCH3), 4.27 (2H, s, NCH2), 4.85 (2H, s, NCH2), 7.14–7.24 (4H, m, aromatics). HRMS (FAB+) calculated for C20H28N3O5: [M+H]+: 390.202896. found: 390.203567.
Reaction of 2-(2,2-dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-methyl-N-(4-trifluoromethylbenzyl)-acetamide (0.50 g, 1.45 mmol) with the paraformaldehyde-methylamine adduct in methanol using a procedure similar to the one described in the preparation of compound 44 (method 44B) gave 0.31 g (64% yield) of the title compound as white crystals; mp 126–128° C. (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ (ppm); 3.07 (3H, s, NCH3), 3.13 (3H, s, NCH3), 4.18 (2H, s, NCH2), 4.74 (2H, s, NCH2), 7.41 (2H, d, J=8.1 Hz, aromatics), 7.64 (2H, d, J=8.1 Hz, aromatics), 10.56 (1H, broad, OH). Anal. Calcd for C15H15F3N2O3: C, 54.88; H, 4.61; N, 8.53. Found: C, 54.93; H, 4.57; N, 8.44.
Reaction of 2-(2,2-dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-methyl-N-(4-trifluoromethyl-benzyl)-acetamide (0.36 g, 1.05 mmol) with the paraformaldehyde-N-(2-aminoethyl)morpholine adduct in methanol using a procedure similar to the one described in the preparation of compound 13 gave 0.16 g (35% yield) of the title compound as a solid after chromatography on reversed phase silica gel. 1HNMR 400 MHz (CDCl3) δ (ppm); 2.46 (4H, m, 2×NCH2), 2.57 (2H, t, J=6.1 Hz, NCH2), 3.04 (3H, s, NCH3), 3.6 (6H, m, 2×OCH2 and NCH2), 4.25 (2H, s, NCH2), 4.72 (2H, s, NCH2), 7.41 (2H, d, J=8.0 Hz, aromatics), 7.62 (2H, d, J=8.0 Hz, aromatics). HRMS (FAB+) calculated for C20H25F3N3O4: [M+H]+: 428.179716. found: 428.179157.
Reaction of 2-(2,2-dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-methoxy-N-(4-trifluoromethylbenzyl)-acetamide (0.20 g, 0.56 mmol) with the paraformaldehyde-methylamine adduct in methanol using a procedure similar to the one described in the preparation of compound 44 (method 44B) gave 0.10 g (52% yield) of the title compound as white crystals; mp 145° C. (dec) (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ (ppm); 3.11 (3H, s, NCH3), 3.75 (3H, s, OCH3), 4.17 (2H, s, NCH2), 4.94 (2H, s, NCH2), 7.45 (2H, d, J=8.1 Hz, aromatics), 7.61 (2H, d, J=8.1 Hz, aromatics). Anal. Calcd for C15H15F3N2O4: C, 52.33; H, 4.39; N, 8.13. Found: C, 52.17; H, 4.34; N, 7.98.
Reaction of 2-(2,2-dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-methoxy-N-(4-trifluoromethyl-benzyl)-acetamide (0.41 g, 1.13 mmol) with the paraformaldehyde-N-(2-aminoethyl)morpholine adduct in methanol using a procedure similar to the one described in the preparation of compound 13 gave 0.38 g (75% yield) of the title compound as crystals after chromatography on reversed phase silica gel; mp 119° C. (dec) (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ (ppm); 2.48 (4H, m, 2×NCH2), 2.58 (2H, t, J=6.3 Hz, NCH2), 3.64 (2H, t, J=6.3 Hz, NCH2), 3.67 (4H, m, 2×OCH2), 3.75 (3H, s, OCH3), 4.31 (2H, s, NCH2), 4.94 (2H, s, NCH2), 7.47 (2H, d, J=8.1 Hz, aromatics), 7.63 (2H, d, J=8.1 Hz, aromatics). Anal. Calcd for C20H24F3N3O5: C, 54.17; H, 5.46; N, 9.48. Found: C, 54.12; H, 5.57; N, 9.52.
Reaction 4-({[2-(2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl]-methoxy-amino}-methyl)-N-methyl-benzamide (0.053 g, 0.15 mmol) with the paraformaldehyde-N-(2-aminoethyl)morpholine adduct in methanol using a procedure similar to the one described in the preparation of compound 13 gave 0.034 g (41% yield) of the title compound as a white solid after chromatography on reversed phase silica gel. 1HNMR 400 MHz (DMSO-d6) δ (ppm); (TFA salt) 2.77 (3H, d, J=4.65 Hz, NCH3), 3.1 (2H, broad m, NCH2), 3.4–3.8 (8H, broad m, 2×NCH2 and 2×OCH2), 3.75 (3H, s, OCH2), 3.98 (2H, broad, NCH2), 4.24 (2H, s, NCH2), 4.94 (2H, s, NCH2), 7.39 (2H, d, J=8.4 Hz, aromatics), 7.80 (2H, d, J=8.4 Hz, aromatic), 8.41 (1H, q, J=4.5 Hz, NH). HRMS (FAB+) calculated for C21H29N4O6: [M+H]+: 433.208710. found: 433.209419.
Reaction of N-(3,4-dichlorobenzyl)-2-(2,2-dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-methoxy-acetamide (0.20 g, 0.56 mmol) with the paraformaldehyde-N-(2-aminoethyl)morpholine adduct in methanol using a procedure similar to the one described in the preparation of compound 13 gave 0.071 g (28% yield) of the title compound as a solid after chromatography on reversed phase silica gel. 1HNMR 400 MHz (DMSO-d6) δ (ppm); 2.39 (4H, broad, 2×NCH2), 2.47 (2H, t, J=6.3 Hz, NCH2), 3.49 (2H, t, J=6.3 Hz, NCH2), 3.53 (4H, broad, 2×OCH2), 3.66 (3H, s, OCH3), 4.16 (2H, s, NCH2), 4.89 (2H, s, NCH2), 7.33 (1H, dd, J=1.8 Hz and J=8.1 Hz, aromatic), 7.60 (1H, d, J=8.1 Hz, aromatic), 7.63 (1H, d, J=1.8 Hz, aromatic). HRMS (MAB/N2) calculated for C19H23C2N3O5: [M+]: 443.101477. found: 443.103002.
Reaction of 2-(2,2-dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-(4-fluoro-2-trifluoromethylbenzyl)-N-methoxy-acetamide (0.14 g, 0.37 mmol) with the paraformaldehyde-methylamine adduct in methanol using a procedure similar to the one described in the preparation of compound 44 (method 44B) gave 0.056 g (42% yield) of the title compound as white crystals; mp 167° C. (dec) (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ (ppm); 3.13 (3H, s, NCH3), 3.71 (3H, s, OCH3), 4.21 (2H, s, NCH2), 5.09 (2H, s, NCH2), 7.22–7.27 (1H, m, aromatic), 7.39–7.46 (2H, m, aromatics). Anal. Calcd for C15H14F4N2O4: C, 49.73; H, 3.89; N, 7.73. Found: C, 49.74; H, 3.92; N, 7.70.
Reaction of 2-(2,2-dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-(4-fluoro-2-trifluoromethyl-benzyl)-N-methoxy-acetamide (0.21 g, 0.55 mmol) with the paraformaldehyde-N-(2-aminoethyl)morpholine adduct in methanol using a procedure similar to the one described in the preparation of compound 13 gave 0.13 g (50% yield) of the title compound as a solid after chromatography on reversed phase silica gel. 1HNMR 400 MHz (CDCl3) δ (ppm); 2.51 (4H, m, 2×NCH2), 2.60 (2H, t, J=6.1 Hz, NCH2), 3.65 (6H, m, 2×OCH2 and NCH2), 3.71 (3H, s, OCH3), 4.32 (2H, s, NCH2), 5.09 (2H, s, NCH2), 7.23–7.27 (2H, m, aromatics), 7.39–7.48 (2H, m, aromatics). MS (ESI+) calculated for C20H24F4N3O5: [M+H]+: 462. found: 462.
Reaction of N-[3-(3,4-dichlorophenyl)-propyl]-2-(2,2-dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-acetamide (0.432 g, 1.20 mmol) with the paraformaldehyde-methylamine adduct in methanol using a procedure similar to the one described in the preparation of compound 44 (method 44B) gave 0.232 g (56% yield) of the title compound as white crystals; mp 157–158° C. (dec) (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ (ppm); 1.81 (2H, m, CH2), 2.57 (2H, t, J=7.5 Hz, CH2), 3.05 (3H, s, NCH3), 3.33 (2H, m, NCH2), 3.98 (2H, s, NCH2), 6.95 (1H, broad t, NH), 7.05 (1H, broad dd, aromatic), 7.21 (1H, d, J=2 Hz, aromatic), 7.26 (1H, d, J=8.1 Hz, aromatic), 10.13 (1H, broad, OH). Anal. Calcd for C5H16Cl2N2O3: C, 52.49; H, 4.70; N, 8.16. Found: C, 52.39; H, 4.80; N, 7.89.
Reaction of 2-(2,2-dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-[3-(4-fluorophenyl)-propyl]-N-methyl-acetamide (0.20 g, 0.62 mmol) with the paraformaldehyde-methylamine adduct in methanol using a procedure similar to the one described in the preparation of compound 44 (method 44B) gave 0.11 g (56% yield) of the title compound as white crystals; mp 129° C. (dec) (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ (ppm); 1.91 (2H, m, CH2), 2.63 (2H, t, J=7.6 Hz, CH2), 3.04 (3H, s, NCH3), 3.05 (3H, s, NCH3), 3.40 (2H, broad t, J=7.5 Hz, NCH2), 3.90 (2H, broad s, NCH2), 6.99 (2H, m, aromatics), 7.14 (2H, m, aromatics). Anal. Calcd for C16H19FN2O3: C, 62.73; H, 6.25; N, 9.14. Found: C, 62.75; H, 6.23; N, 9.11.
Reaction of 2-(2,2-dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-[3-(4-fluorophenyl)-propyl]-N-methyl-acetamide (0.245 g, 0.76 mmol) with the paraformaldehyde —N-(2-aminoethyl)morpholine adduct in methanol using a procedure similar to the one described in the preparation of compound 13 gave 0.103 g (33% yield) of the title compound as a solid after chromatography on reversed phase silica gel. 1HNMR 400 MHz (CDCl3) δ (ppm); 1.92 (2H, m, CH2), 2.47 (4H, broad, 2×NCH2), 2.54 (2H, broad t, NCH2), 2.63 (2H, t, J=7.5 Hz, CH2), 3.04 (3H, s, NCH3), 3.42 (2H, broad t, NCH2), 3.58 (2H, broad, NCH2), 3.66 (4H, broad, 2×OCH2), 4.12 (2H, broad s, NCH2), 6.97 (2H, m, aromatics), 7.14 (2H, m, aromatics). HRMS (FAB+) calculated for C21H29FN3O4: [M+H]+: 406.214210. found: 406.214016.
Reaction of N-[3-(3,4-dichlorophenyl)-propyl]-2-(2,2-dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-methyl-acetamide (0.570 g, 1.53 mmol) with the paraformaldehyde-methylamine adduct in methanol using a procedure similar to the one described in the preparation of compound 44 (method 44B) gave 0.287 g (52% yield) of the title compound as white crystals; mp 138–140° C. (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ (ppm); 1.80 (2H, m, CH2), 2.49 (2H, t, J=7.6 Hz, CH2), 2.92 (3H, s, NCH3), 2.95 (3H, s, NCH3), 3.31 (2H, broad t, J=7.3 Hz, NCH2), 3.87 (2H, broad s, NCH2), 6.91 (1H, dd, J=2.0 Hz and J=8.1 Hz, aromatic), 7.16 (1H, d, J=2.0 Hz, aromatic), 7.23 (1H, d, J=8.1 Hz, aromatic), 10.94 (1H, broad s, OH). Anal. Calcd for C6H18Cl2N2O3: C, 53.80; H, 5.08; N, 7.84. Found: C, 53.90; H, 5.17; N, 7.83.
Reaction of 2-(2,2-dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-[3-(4-fluorophenyl)-propyl]-N-methoxy-acetamide (0.560 g, 1.66 mmol) with the paraformaldehyde-N-(2-aminoethyl)morpholine adduct in methanol using a procedure similar to the one described in the preparation of compound 13 gave 0.446 g (63% yield) of the title compound as a solid after chromatography on reversed phase silica gel. 1HNMR 400 MHz (DMSO-d6) δ (ppm); 1.84 (2H, m, CH2), 2.40 (4H, broad, 2×NCH2), 2.46 (2H, t, J=6.5 Hz, CH2), 2.56 (2H, t, J=8.1 Hz, CH2), 3.46 (2H, t, J=6.5 Hz, NCH2), 3.52 (3H, s, OCH3), 3.55 (4H, broad, 2×OCH2), 3.67 (2H, t, J=7.1 Hz, NCH2), 3.95 (2H, s, NCH2), 7.09 (2H, m, aromatics), 7.23 (2H, m, aromatics). HRMS (FAB+) calculated for C21H29FN3O5: [M+H]+: 422.209125. found: 422.208679.
Reaction of N-[3-(3,4-dichlorophenyl)-propyl]-2-(2,2-dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-methoxy-acetamide (0.480 g, 1.16 mmol) with the paraformaldehyde-methylamine adduct in methanol using a procedure similar to the one described in the preparation of compound 44 (method 44B) gave 0.250 g (58% yield) of the title compound as white crystals; mp 106–108° C. (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ (ppm); 2.01 (2H, m, CH2), 2.65 (2H, t, J=7.6 Hz, CH2), 3.14 (3H, s, NCH3), 3.73 (3H, s, OCH3), 3.77 (2H, t, J=7.0 Hz, NCH2), 4.11 (2H, s, NCH2), 7.04 (1H, dd, J=2.0 Hz and J=8.1 Hz, aromatic), 7.30 (1H, d, J=2.0 Hz, aromatic), 7.36 (1H, d, J=8.1 Hz, aromatic), 11.67 (1H, broad s, OH). Anal. Calcd for C16H18Cl2N2O4: C, 51.49; H, 4.86; N, 7.51. Found: C, 51.65; H, 4.90; N, 7.45.
Reaction of N-[3-(3,4-dichlorophenyl)-propyl]-2-(2,2-dimethyl-5-oxo-[1,3]-dioxolan-4-ylidene)-N-methoxy-acetamide (0.378 g, 0.97 mmol) with the paraformaldehyde-N-(2-aminoethyl)morpholine adduct in methanol using a procedure similar to the one described in the preparation of compound 13 gave 0.196 g (42% yield) of the title compound as a solid after chromatography on reversed phase silica gel. 1HNMR 400 MHz (CDCl3) δ (ppm); 1.98 (2H, m, CH2), 2.49 (4H, broad, 2×NCH2), 2.58 (2H, t, J=6.2 Hz, CH2), 2.63 (2H, t, J=7.5 Hz, CH2), 3.63 (2H, t, J=6.2 Hz, NCH2), 3.68 (4H, broad, 2×OCH2), 3.71 (3H, s, OCH3), 3.74 (2H, t, J=7.0 Hz, NCH2), 4.23 (2H, s, NCH2), 7.02 (1H, dd, J=2.0 Hz and J=8.2 Hz, aromatic), 7.28 (1H, d, J=2.0 Hz, aromatic), 7.34 (1H, d, J=8.2 Hz, aromatic). HRMS (FAB+) calculated for C21H28Cl2N3O5: [M+H]+: 472.140602. found: 472.138651.
A suspension of sodium hydride (73.0 mmol, 3.0 g of a 60% suspension in mineral oil) was washed twice with hexane and then suspended in dry tetrahydrofuran (40 ml). The reaction mixture was then treated at 25° C. with 1-bromo-3-chloropropane (10 ml, 97.5 mmol) followed by a solution of 4-fluorobenzaldehyde oxime (6.78 g, 48.7 mmol) in tetrahydrofuran (30 ml) added dropwise over 10 min. The resulting mixture was then heated under reflux for 16 h. The cooled mixture was diluted with ethyl acetate, washed with brine and dried over anhydrous sodium sulfate. Evaporation of the solvent under reduced pressure and chromatography of the residue on silica gel (elution hexane-ethyl acetate, 9:1) gave 8.52 g (81%) of the title oxime as a clear oil. 1HNMR 400 MHz (CDCl3) δ (ppm): 2.23 (2H, m, CH2), 3.74 (2H, t, J=6.5 Hz, CH2), 4.36 (2H, t, J=5.8 Hz, CH2), 7.13 (2H, m, aromatics), 7.63 (2H, m, aromatics), 8.11 (1H, s, CH). Anal. Calcd for C10H11ClFNO: C, 55.69; H, 5.14; N, 6.49. Found: C, 55.44; H, 5.12; N, 6.41.
A mixture of 4-fluorobenzaldehyde O-(3-chloropropyl)-oxime (0.430 g, 2.0 mmol), morpholine (0.70 g, 8.0 mmol), sodium iodide (0.1 g) and potassium carbonate (0.55 g) in acetone (10 ml) was sealed and heated at 80° C. for 34 h. The cooled mixture was concentrated, diluted with ethyl acetate, washed with brine and dried over anhydrous sodium sulfate. Evaporation of the solvent under reduced pressure and chromatography of the residue on silica gel (elution ethyl acetate-acetone, 8:2) gave 0.376 g (70%) of the title oxime as a clear oil. 1HNMR 400 MHz (C6D6) δ (ppm): 1.94 (2H, m, CH2), 2.27 (4H, m, NCH2), 2.38 (2H, t, J=7.1 Hz, NCH2), 3.69 (4H, m, OCH2), 4.40 (2H, t, J=6.5 Hz, OCH2), 6.76 (2H, m, aromatics), 7.33 (2H, m, aromatics), 8.03 (1H, s, CH).
Reduction of 4-fluorobenzaldehyde O-(3-morpholin-4-yl-propyl)-oxime (0.330 g, 1.24 mmol) with sodium cyanoborohydride in acetic acid as described in the preparation of compound 44-B gave 0.330 (100% yield) of the crude title hydroxylamine as a light yellow oil which was used as such for the acylation step. 1HNMR 400 MHz (C6D6) δ (ppm): 1.69 (2H, m, CH2), 2.22 (4H, m, NCH2), 2.27 (2H, t, J=7.0 Hz, NCH2), 3.63 (4H, m, OCH2), 3.74 (2H, t, J=6.3 Hz, OCH2), 3.79 (2H, s, NCH2), 6.91 (2H, m, aromatics), 7.12 (2H, m, aromatics).
Reaction of (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl chloride (1.0 mmol) with N-(4-fluorobenzyl)-O-(3-morpholin-4-yl-propyl)-hydroxylamine (0.268 g, mmol) as described in the preparation of compound 44-C (acid wash was skipped in this case) gave 0.200 g (47% yield) of the title amide as a clear oil after chromatography on silica gel. 1HNMR 400 MHz (C6D6) δ (ppm): 1.10 (6H, s, CH3), 1.50 (2H, m, CH2), 2.19 (2H, t, J=6.6 Hz, NCH2), 2.20 (4H, m, NCH2), 3.70 (2H, t, J=6.2 Hz, OCH2), 3.71 (4H, m, OCH2), 4.73 (2H, s, NCH2), 6.85 (3H, m, CH and aromatics), 7.22 (2H, m, aromatics). HRMS (MAB N2) calculated for C21H27FN2O6 [M+]: 422.185315: found: 422.185246.
Reaction of 2-(2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-(4-fluoro-benzyl)-N-(3-morpholin-4-yl-propoxy)-acetamide (0.080 g, 0.18 mmol) with the paraformaldehyde-methylamine adduct in methanol using a procedure similar to the one described in the preparation of compound 44 method 44B) gave 0.027 g (35% yield) of the title compound as a white solid after chromatography on reversed phase silica gel. 1HNMR 400 MHz (DMSO-d6) δ (ppm); 1.61 (2H, m, CH2), 2.23 (2H, t, J=7.0 Hz, NCH2), 2.28 (4H, broad, 2×NCH2), 2.93 (3H, s, NCH3), 3.54 (4H, broad, 2×OCH2), 3.76 (2H, broad t, OCH2), 3.97 (2H, s, NCH2), 4.88 (2H, s, NCH2), 7.13 (2H, m, aromatics), 7.40 (2H, m, aromatics). HRMS (MAB N2) calculated for C20H26FN3O5: [M]+: 407.185650. found: 407.184331.
Reaction of 2-(2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-(4-fluoro-benzyl)-N-(3-morpholin-4-yl-propoxy)-acetamide (0.080 g, 0.19 mmol) with the paraformaldehyde-N-(2-aminoethyl)morpholine adduct in methanol using a procedure similar to the one described in the preparation of compound 13 gave 0.019 g (20% yield) of the title compound as a solid after chromatography on reversed phase silica gel. 1HNMR 400 MHz (CDCl3) δ (ppm); 1.65 (2H, m, CH2), 2.25 (2H, t, J=7.3 Hz, NCH2), 2.29 (4H, broad, 2×NCH2), 2.39 (4H, broad, 2×NCH2), 2.46 (2H, t, J=6.6 Hz, CH2), 3.50 (2H, t, J=6.6 Hz, NCH2), 3.68 (8H, broad, 4×OCH2), 3.84 (2H, t, J=7.0 Hz, OCH2), 4.11 (2H, s, NCH2), 4.87 (2H, s, NCH2), 7.16 (2H, m, aromatics), 7.40 (2H, m, aromatics). HRMS (MAB N2) calculated for C25H35FN4O6: [M]+: 506.254064; found: 506.254892.
Reaction of 3,4-dichlorobenzaldehyde oxime (2.5 g, 13.15 mmol) with 1-bromo-3-chloropropane (2.6 ml, 26.3 mmol) as described in the preparation of Compound 146-A gave 2.60 g (74% yield) of the title oxime as a clear oil after chromatography on silica gel (elution hexane-ethyl acetate, 9:1). 1HNMR 400 MHz (CDCl3) δ (ppm): 2.21 (2H, m, CH2), 3.69 (2H, t, J=6.5 Hz, CH2), 4.34 (2H, t, J=5.8 Hz, OCH2), 7.41 (1H, dd, J=2 Hz and J=8 Hz, aromatic), 7.47 (1H, d, J=8 Hz, aromatic), 7.71 (1H, d, J=2 Hz, aromatic), 8.01 (1H, s, CH).
Reaction of 3,4-dichlorobenzaldehyde O-(3-chloropropyl)-oxime (2.6 g, 9.75 mmol) with morpholine (3.4 ml) as described in the preparation of compound 146-B gave 2.25 g (72% yield) the title oxime as a clear oil after chromatography on silica gel (elution ethyl acetate-acetone, 8:2). 1HNMR 400 MHz (C6D6) δ (ppm): 1.89 (2H, m, CH2), 2.25 (4H, m, NCH2), 2.35 (2H, t, J=7.1 Hz, NCH2), 3.69 (4H, m, OCH2), 4.35 (2H, t, J=6.5 Hz, OCH2), 6.98 (1H, d, J=8.5 Hz, aromatic), 7.02 (1H, dd, J=1.9 Hz and J=8.5 Hz, aromatic), 7.50 (1H, d, J=1.9 Hz, aromatic), 7.77 (1H, s, CH). Anal. Calcd for C14H18Cl2N2O2: C, 53.01; H, 5.71; N, 8.83. Found: C, 52.99; H, 5.69; N, 8.75.
A solution of 3,4-dichlorobenzaldehyde O-(3-morpholin-4-yl-propyl)-oxime (0.160 g, 0.5 mmol) in a mixture of dichloromethane (2 ml) and acetic acid (1 ml) was treated with borane-pyridine complex (0.17 ml, 1.36 mmol) and the resulting mixture was heated under reflux (bath temperature 60° C.) for 4 h. The solvent was then evaporated under reduced pressure and the residue was treated with 10% aqueous hydrochloric acid (3 ml). The reaction mixture was then basified with solid sodium carbonate and extracted with ethyl acetate. The organic phase was then washed with brine, dried (anhydrous magnesium sulfate) and concentrated to give 0.150 g (93% yield) of the crude title hydroxylamine as a light yellow oil which was used as such for the acylation step. 1HNMR 400 MHz (C6D6) δ (ppm): 1.66 (2H, m, CH2), 2.22 (2H, t, J=7.1 Hz, NCH2), 2.24 (4H, m, NCH2), 3.56 (2H, s, NCH2), 3.69 (4H, m, OCH2), 3.70 (2H, t, J=6.5 Hz, OCH2), 5.2 (1H, broad, NH), 6.82 (1H, dd, J=1.9 Hz and J=8.5 Hz, aromatic), 7.13 (1H, d, J=8.5 Hz, aromatic), 7.33 (1H, d, J=1.9 Hz, aromatic).
Reaction of (2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-acetyl chloride (0.49 mmol) with N-(3,4-dichlorobenzyl)-O-(3-morpholin-4-yl-propyl)-hydroxylamine (0.150 g, 0.49 mmol) as described in the preparation of compound 44-C gave 0.150 g (65% yield) of the title amide as white crystals after chromatography on silica gel: mp 77–78° C. (ether-hexane). 1HNMR 400 MHz (C6D6) δ (ppm): 1.09 (6H, s, CH3), 1.47 (2H, m, CH2), 2.17 (2H, t, J=6.5 Hz, NCH2), 2.22 (4H, m, NCH2), 3.66 (2H, t, J=6.0 Hz, OCH2), 3.73 (4H, m, OCH2), 4.56 (2H, s, NCH2), 6.82 (1H, s, CH), 6.98 (1H, dd, J=2.0 Hz and J=8.0 Hz, aromatic), 7.06 (1H, d, J=8.0 Hz, aromatic), 7.40 (1H, d, J=2.0 Hz, aromatic). Anal. Calcd for C21H26Cl2N2O6: C, 53.28; H, 5.53; N, 5.91. Found: C, 53.31; H, 5.67; N, 5.77.
Reaction of N-(3,4-dichloro-benzyl)-2-(2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-(3-morpholin-4-yl-propoxy)-acetamide (0.200 g, 0.42 mmol) with the paraformaldehyde-methylamine adduct in methanol using a procedure similar to the one described in the preparation of compound 44 (method 44B) gave 0.090 g (47% yield) of the title compound as a white amorphous solid after chromatography on reversed phase silica gel. 1HNMR 400 MHz (DMSO-d6) δ (ppm); 1.66 (2H, m, CH2), 2.27 (2H, t, J=7.1 Hz, NCH2), 2.32 (4H, broad, 2×NCH2), 2.95 (3H, s, NCH3), 3.55 (4H, broad, 2×OCH2), 3.86 (2H, broad t, OCH2), 4.06 (2H, s, NCH2), 4.88 (2H, s, NCH2), 7.35 (1H, dd, J=2 Hz and J=8.1 Hz, aromatic), 7.61 (1H, d, J=8.1 Hz, aromatic), 7.65 (1H, d, J=2 Hz, aromatic). HRMS (FAB+) calculated for C20H26Cl2N3O5: [M+H]+: 458.124952. found: 458.123753.
Compound 875: 4-Hydroxy-1-(2-morpholin-4-yl-ethyl)-5-oxo-2,5-dihydro-1H-pyrrole-3-carboxylic acid (3,4-dichloro-benzyl)-(3-morpholin-4-yl-propoxy)-amide
Reaction of N-(3,4-dichloro-benzyl)-2-(2,2-dimethyl-5-oxo-[1,3]dioxolan-4-ylidene)-N-(3-morpholin-4-yl-propoxy)-acetamide (0.200 g, 0.42 mmol) with the paraformaldehyde-N-(2-aminoethyl)morpholine adduct in methanol using a procedure similar to the one described in the preparation of compound 13 gave 0.160 g (68% yield) of the title compound as a solid after chromatography on reversed phase silica gel. 1HNMR 400 MHz (CDCl3) δ (ppm); 1.56 (2H, m, CH2), 2.18 (2H, t, J=7.1 Hz, NCH2), 2.25 (4H, broad, 2×NCH2), 2.39 (4H, broad, 2×NCH2), 2.46 (2H, t, J=6.1 Hz, CH2), 3.50 (2H, t, J=6.1 Hz, NCH2), 3.54 (8H, broad, 4×OCH2), 3.71 (2H, broad t, OCH2), 3.99 (2H, s, NCH2), 4.87 (2H, s, NCH2), 7.40 (1H, broad d, aromatic), 7.58 (1H, d, J=8.1 Hz, aromatic), 7.73 (1H, broad s, aromatic). HRMS (FAB+) calculated for C25H35Cl2N4O6: [M+H]+: 557.193366. found: 557.192134.
For each reaction, 5 pmole of biotin labeled substrate DNA was bound to 100 ug of Streptavidin coated PVT SPA beads (Amersham Pharmacia Biotech). 0.26 ng of recombinant integrase was incubated with the beads for 90 min at 37 C. Unbound enzyme was removed by washing the complex followed by addition of inhibitors and 0.1 fmol of P33 labeled target DNA. Reaction was stopped by adding EDTA to a final concentration of 10 mM. Samples were counted in TopCountNXT (Packard) and the CPM was used as a measure of integration. Reaction condition was as described in A. Engelman and R. Craigie, J. Virol. 69, 5908–5911 (1995). The sequences of substrate and target DNA were described in Nucleic Acid Research 22, 1121–1122 (1994). Using this assay, representative examples were found to have IC50=0.01 to 50 μM. The table below shows the percent inhibition of HIV-integrase in the presence of 20 μM of compounds 1–79.
Inhibition of HIV Replication
Cell culture assays were performed using a single cycle, recombinant HIV virus expressing Renella luciferase. Anti-viral activity was evaluated by measuring the production of luciferase in the infected cells 5 days post-infection. Susceptibility of the virus to compounds was determined by incubation in the presence of the serially-diluted compound. The 50% effective concentration (EC50) was calculated by using the exponential form of the median effect equation where (Fa)=1/[1+(ED50/drug conc.)m].
Representative compounds of this invention tested in this assay have EC50's of approximately 0.01 to 150 μM.
This application claims the benefit of U.S. Provisional Application No. 60/399,248, filed Jul. 29, 2002 and U.S. Provisional Application No. 60/394,548, filed Jul. 9, 2002.
| Number | Name | Date | Kind |
|---|---|---|---|
| 6727275 | Zou et al. | Apr 2004 | B1 |
| Number | Date | Country |
|---|---|---|
| WO 02070491 | Sep 2002 | WO |
| WO 03016266 | Feb 2003 | WO |
| WO 03016275 | Feb 2003 | WO |
| WO 03035076 | May 2003 | WO |
| WO 03035077 | May 2003 | WO |
| Number | Date | Country | |
|---|---|---|---|
| 20040110804 A1 | Jun 2004 | US |
| Number | Date | Country | |
|---|---|---|---|
| 60399248 | Jul 2002 | US | |
| 60394548 | Jul 2002 | US |
