Patent Application
20060211755
This invention relates to novel dihydro-1H-indazole-5-carboxylic acid derivative compounds useful for the treatment and/or prevention of diseases mediated by phospholipases including hepatic lipase and endothelial lipase.
Hepatic lipase plays an important role in lipid metabolism. Hepatic lipase is a glycoprotein that functions as a ligand or as an enzyme of approximately 65 Kda, which has been shown to catalyze the hydrolysis of lipids including triglycerides, diglycerides and phospholipids in native lipoproteins. It has also been shown to facilitate the selective uptake of cholesterol from high-density lipoproteins and the removal of remnant particles by the liver (Jonathan C. Cohen, et al Biochemistry 1992, 31: 8544-8551 and Neve et al Biochemistry J. 1998, 330:701-706).
Other studies showing the inverse relationship of HDL and hepatic lipase activity include for example, Haffner S. M. et al., “Studies on the metabolic mechanism of reduced high density lipoproteins during anabolic steroid therapy,” Metabolism 1983; 32:413-420; Applebaum-Bowden D, et al., “The Dyslipoproteinemia of Anabolic steroid therapy: increase in hepatic triglyceride lipase precedes the decrease in high density lipoprotein −2 cholesterol,” Metabolism 1987; 36:949-952; and Kantor M. A. et al., “Androgens reduce HDL-2 cholesterol and increase hepatic triglyceride lipase activity,” Med. Sci. Sport exercise 1985; 17:462-465.
The inverse relationship between hepatic lipase activity and the level of HDL-cholesterol, particularly type-2 HDL-cholesterol, can be used to advantage in up-regulating the Level of HDL cholesterol-the good cholesterol.
Endothelial lipase (EL) is a newly described member of the lipase gene family. Like hepatic lipase, endothelial lipase has been implicated in the hydrolysis of HDL phospholipids and in the reduction of HDL-cholesterol in vivo.
In experiments using hepatic lipase knockout mice the infusion of a polyclonal antibody inhibitory to endothelial lipase resulted in a marked increase in HDL-cholesterol levels (Rader, D. J., et al Journal of Clinical Investigation (2003), 111(3) 357-362.
Chan, et al, Proceedings of the National Academy of Sciences U.S.A. (2003), 100(5), 2748-2753, has also reported the inverse relation between endothelial lipase and HDL-cholesterol.
Given the preceding information, it is desirable to discover and develop compounds that increase HDL levels by methods that may include inhibiting the activity of hepatic lipase and/or endothelial lipase in order to treat, prevent and/or ameliorate the effects of hepatic lipase and/or endothelial lipase mediated diseases. Few therapeutically desirable agents are available to accomplish the task of increasing HDL levels hence the need for and utility of the present invention.
The present invention provides a compound of formula I
wherein;
R1 is selected from the group consisting of C5-C13alkyl, C1-C12haloalkyl, C4-C12alkenyl, C4-C12alkynyl, or C1-C5alkylcycloalkyl, C3-C8cycloalkyl, C1-C5alkylheterocyclic, and aryl, wherein the, cycloalkyl, cycloalkenyl, heterocyclic and aryl substituents are optionally substituted with one to three substituents independently selected from C1-C8alkyl, C1-C8haloalkyl, C2-C8alkenyl, C2-C8alkynyl, phenyl, benzyl, hydroxy, C1-C5 alkoxy, (CH2)mCOOC1-C5alkyl, (CH2)mNRaRb, and C1-C4alkylcycloalkyl; wherein Ra and Rb are independently selected from hydrogen, C1-C8alkyl, C2-C8alkenyl, C2-C8alkynyl, and C1-C5alkylcycloalkyl;
R2 is hydrogen;
R3, R4, R5, and R6, are independently selected from hydrogen, C1-C12alkyl, C2-C12haloalkyl, C2-C12alkenyl, C2-C12alkynyl, C2-C12alkylaryl, C1-C12alkylcyclohexyl, C1-C12alkylcyclopentyl, C1-C12alkylheterocyclic, (CH2)mCOOH, (CH2)mCO(C1-C10)alkyl, (CH2)mCOO(C1-C10)alkyl, (CH2)mCOO(C1-C10)alkylaryl, C1-C10alkylamino, halo, (CH2)mCONH2, (CH2)mCONRaRb, phenyl, or aryl wherein each of the phenyl or aryl groups is optionally substituted with one to three groups independently selected from C1-C8alkyl, C1-C8haloalkyl, C2-C8alkenyl, C2-C8alkynyl, phenyl, benzyl, hydroxy, C1-C5 alkoxy, (CH2)mCOOC1-C5alkyl, and C1-C4alkylcycloalkyl; and wherein m is 0, 1, 2, or 3;
R7 is selected from the group consisting of hydrogen, C1-C10alkyl, C1-C10haloalkyl, C2-C10alkenyl, C2-C10alkynyl, C1-C6alkylaryl, C1-C6alkylcyclohexyl, C1-C6alkylcyclopentyl, C1-C6alkylheterocyclic, or aryl; or a pharmaceutically acceptable salt, solvate or isomer thereof.
The present invention relates to the use of a compound of formula (I)
or a pharmaceutically acceptable salt, solvate or enantiomer thereof, for the treatment, amelioration and/or prevention of diseases mediated by hepatic lipase and/or endothelial lipase activity:
wherein;
R1 is selected from the group consisting of C5-C13alkyl, C1-C12haloalkyl, C4-C12alkenyl, C4-C12alkynyl, or C1-C5alkylcycloalkyl, C3-C8cycloalkyl, C1-C5alkylheterocyclic, and aryl, wherein the, cycloalkyl, cycloalkenyl, heterocyclic and aryl substituents are optionally substituted with one to three substituents independently selected from C1-C8alkyl, C1-C8haloalkyl, C2-C8alkenyl, C2-C8alkynyl, phenyl, benzyl, hydroxy, C1-C5 alkoxy, (CH2)mCOOC1-C5alkyl, (CH2)mNRaRb, and C1-C4alkylcycloalkyl; wherein Ra and Rb are independently selected from hydrogen, C1-C8alkyl, C2-C8alkenyl, C2-C8alkynyl, and C1-C5alkylcycloalkyl;
R2 is hydrogen;
R3, R4, R5, and R6, are independently selected from hydrogen, C1-C12alkyl, C2-C12haloalkyl, C2-C12alkenyl, C2-C12alkynyl, C2-C12alkylaryl, C1-C12alkylcyclohexyl, C1-C12alkylcyclopentyl, C1-C12alkylheterocyclic, (CH2)mCOOH, (CH2)mCO(C1-C10)alkyl, (CH2)mCOO(C1-C10)alkyl, (CH2)mCOO(C1-C10)alkylaryl, C1-C10alkylamino, halo, (CH2)mCONH2, (CH2)mCONRaRb, phenyl, or aryl wherein each of the phenyl or aryl groups is optionally substituted with one to three groups independently selected from C1-C8alkyl, C1-C8haloalkyl, C2-C8alkenyl, C2-C8alkynyl, phenyl, benzyl, hydroxy, C1-C5 alkoxy, (CH2)mCOOC1-C5alkyl, and C1-C4alkylcycloalkyl; and wherein m is 0, 1, 2, or 3;
R7 is selected from the group consisting of hydrogen, C1-C10alkyl, C1-C10haloalkyl, C2-C10alkenyl, C2-C10alkynyl, C1-C6alkylaryl, C1-C6alkylcyclohexyl, C1-C6alkylcyclopentyl, C1-C6alkylheterocyclic, or aryl; or a pharmaceutically acceptable salt, solvate or isomer thereof.
The present invention provides novel dihydro-1H-indazole-5-carboxylic acid derivative compounds of formula I having potent and selective effectiveness as inhibitors of mammalian hepatic lipase and/or endothelial lipase.
The present invention also relates to the use of a novel dihydro-1H-indazole-5-carboxylic acid derivative compound of formula I to increase or mediate the increase of high-density lipoproteins (HDL) upon administration to a patient in need thereof.
The present invention provides a pharmaceutical composition containing any of the compounds of formula I.
The present invention relates to the use of a compound of formula for the treatment and/or prevention or amelioration of diseases related to or exacerbated by hepatic lipase and/or endothelial lipase activity comprising administering the compound of formula I to a patient in need thereof.
The present invention also relates to the use of a pharmaceutical composition comprising a compound of formula I and a carrier and/or diluent for the treatment and/or prevention of hypercholesterolemia.
The present invention relates to the use of a pharmaceutical composition comprising a therapeutically effective amount of hepatic lipase and/or endothelial lipase inhibitor compound of formula I and mixtures thereof for the manufacture of a medicament for the treatment of hepatic lipase and/or endothelial lipase-mediated diseases.
The phrase, “hepatic lipase and/or endothelial lipase mediated-diseases” refers to diseases symptomatic of low HDL levels, caused by, modulated by, exacerbated by or induced directly or indirectly by elevated hepatic lipase and/or endothelial lipase activity, and include for example, hypercholesterolemia; hyperlipidemia, stroke, hypertriglyceridemia, atherosclerosis and related diseases. Treatment and/or prevention of such diseases comprises administering to a mammal in need of such treatment a therapeutically effective amount of the compound of formula I in an amount sufficient to inhibit, ameliorate and/or prevent hepatic lipase and/or endothelial lipase activity and to thereby inhibit or prevent the deleterious effects of hepatic lipase and/or endothelial lipase activity.
The term “Active Ingredient” as used herein refers to a compound(s) of Formula (I) or a pharmaceutically acceptable salt, solvate, prodrug, racemate or enantiomer thereof either as the pure compound or delivered as a pharmaceutical formulation or a pharmaceutical composition. The pharmaceutical composition or formulation containing a compound of the invention and other compound(s) or treatment regimens useful for the treatment and/or prevention of diseases associated with or exacerbated by hepatic lipase and/or endothelial lipase activity (combination drugs) are contemplated to be within the meaning of the term “Active Ingredient(s).”
The term, “indazole nucleus”, or “dihydro-1H-indazole nucleus” as used herein refers to a nucleus (having numbered positions) with the structural formula (X):
The dihydro-1H-indazole derivative compounds of the invention employ certain defining terms as follows:
The term, “alkyl” by itself or as part of another substituent means, unless otherwise defined, a straight or branched chain monovalent hydrocarbon radical such as methyl, ethyl, n-propyl, isopropyl, n-butyl, tertiary butyl, sec-butyl, n-pentyl, and n-hexyl.
The term, “alkenyl” employed alone or in combination with other terms means a straight chain or branched monovalent hydrocarbon group having the stated number ranges of carbon atoms, and typified by groups such as vinyl, propenyl, crotonyl, isopentenyl, and various butenyl isomers.
The term, “hydrocarbyl” means an organic group containing only carbon and hydrogen.
The term, “halo” means fluoro, chloro, bromo, or iodo.
The term, “heterocyclic radical or heterocyclic group”, refers to radicals or groups derived from monocyclic or polycyclic, saturated or unsaturated, substituted or unsubstituted heterocyclic nuclei having 5 to 14 ring atoms and containing from 1 to 3 hetero atoms selected from the group consisting of nitrogen, oxygen or sulfur. Typical heterocyclic radicals are pyrrolyl, pyrrolodinyl, piperidinyl, furanyl, thiophenyl, pyrazolyl, imidazolyl, phenylimidazolyl, triazolyl, isoxazolyl, oxazolyl, thiazolyl, thiadiazolyl, benz(b)thiophenyl, carbazolyl, norharmanyl, azabenzo(b)thiophenyl, benzofuranyl, dibenzofuranyl, dibenzothiophenyl, indazolyl, imidazo(1.2-A)pyridinyl, benzotriazolyl, anthranilyl, 1,2-benzisoxazolyl, benzoxazolyl, benzothiazolyl, purinyl, pyridinyl, dipyridylyl, phenylpyridinyl, benzylpyridinyl, pyrimidinyl, phenylpyrimidinyl, pyrazinyl, 1,3,5-triazinyl, quinolinyl, phthalazinyl, quinazolinyl, morpholino, thiomorpholino, homopiperazinyl, tetrahydrofuranyl, tetrahydropyranyl, oxacanyl, 1,3-dioxolanyl, 1,3-dioxanyl, 1,4-dioxanyl, tetrahydrothiophenyl, pentamethylenesulfadyl, 1,3-dithianyl, 1,4-dithianyl, 1,4-thioxanyl, azetidinyl, hexamethyleneiminium, heptamethyleneiminium, piperazinyl and quinoxalinyl.
The terms “C1-C12alkylcyclopentyl,” “C1-C12alkylcyclohexyl,” or “C1-C12alkylheterocyclic” represent respectively a C1-C12alkyl, C1-C12alkyl, or C1-C12alkyl attached to a cylopentyl, cyclohexyl, and heterocyclic group respectively, wherein the entire group is attached to the dihydro-1H-indazole nucleus (X) or other substrate via the alkyl terminus at indicated or designated positions. The term “cycloalkyl” without more implies a cycloalkyl group having from 3 to 8 carbon atoms.
The term “substituted group” is an organic group substituted with one or more suitable substituents. For example, substituted phenyl as used herein refers to a phenyl group having one to three substituents selected from C1-C12alkyl, C2-C12haloalkyl, C2-C12alkenyl, C2-C12alkynyl, C2-C12alkylaryl, C1-C12alkylcyclohexyl, C1-C12alkylcyclopentyl, C1-C12alkylheterocyclic, (CH2)mCOOH, (CH2)mCO(C1-C10)alkyl, (CH2)mCOO(C1-C10)alkyl, (CH2)mCOO(C1-C10)alkylaryl, C1-C10alkylamino, halo, (CH2)mCONH2, (CH2)mCON((C1-C6)alkyl)2, phenyl, substituted phenyl, or aryl, wherein m=0, 1, 2, or 3. Similarly, the term substituted benzyl means a benzyl group (CH2Phenyl) having substitution on the phenyl ring as described above. Analogously, the term aryl as used herein has its usual meaning and especially refers to the benzyl group.
As used herein the terms “group”, “radical” or “fragment” are synonymous and are intended to indicate functional groups or fragments of molecules attachable to a bond or other fragments of molecules. For example acetamide group represent the acetamide fragment or radical. Structures of groups, radicals or fragments unattached to the dihydro-1-H-indazole-5-carboxylic acid derivative nucleus have been drawn to show the first line as a connecting bond only. Thus, the group
represents the acetamide radical or group, not the propanamide radical unless otherwise indicated.
Preferred R1 Substituents:
The preferred group for R1 is a substituted or unsubstituted group selected from the group consisting of C5-C13alkyl, C1-C5alkylcycloalkyl, C4-C12cycloalkenyl, cycohexylmethyl, cyclopentylmethyl, cyclohexylethyl, substituted or unsubstituted benzyl.
A most preferred R1 is selected from the group consisting of 2, 4 or 5-substituted benzyl, 2,5 disubstituted benzyl, 2,4-disubstituted benzyl, or 3,5-disubstitued benzyl. Preferred substituents on the benzyl group include methyl, ethyl, propyl, butyl, pentyl, hexyl, cyclohexylmethyl, phenyl, and cycloheptylmethyl. Most preferred substituents on the benzyl group include methyl, ethyl, propyl, isopropyl, isobutyl, tert-butyl, and pentane.
Preferred R3, R4, R5, and R6 Substituents:
R3, R4, R5, and R6 are preferably independently selected from the group consisting of hydrogen, C1-C6alkyl, C2-C6alkenyl, —O—(C1-C6)alkyl, —S—(C1-C6)alkyl, —C5-C12cycloalkyl, (CH2)mCOOH, (CH2)mCO(C1-C10)alkyl, (CH2)mCOO(C1-C10)alkyl, (CH2)mCOO(C1-C10)alkylaryl, C1-C10alkylamino, halo, (CH2)mCONH2, (CH2)mCONRaRb, phenyl, substituted phenyl, or aryl. Particularly preferred R3, R4, R5, and R6 groups are selected from hydrogen, methyl, ethyl, propyl, chloro, fluoro, —COOH, CO(C1-C10)alkyl, COO(C1-C10)alkyl, and CONH2 and salts thereof More particularly hydrogen, chloro, and sodium, potassium and lithium salts of —COOH are preferred as independently selected R3, R4, R5, and R6 groups.
A preferred compound of the invention is a compound selected from the group consisting of:
More preferred compounds of the invention are represented by the formulae (C1), (C2), (C3), (C4) and (C5):
The salts of the 1H-indazole compounds represented by formula (I), are an additional aspect of the invention.
In those instances when the compound of the invention possesses acidic or basic functional groups, various salts may be formed which are more water soluble and more physiologically suitable than the parent compound. Representative pharmaceutically acceptable salts, include but are not limited to, the alkali and alkaline earth salts such as lithium, sodium, potassium, calcium, magnesium, aluminum and the like. Salts are conveniently prepared from the free acid by treating the acid in solution with a base or by exposing the acid to an ion-exchange resin.
Included within the definition of pharmaceutically acceptable salts are the relatively non-toxic, inorganic and organic base addition salts of compounds of the present invention, for example, ammonium, quaternary ammonium, and amine cations, derived from nitrogenous bases of sufficient basicity to form salts with the compounds of this invention (see, for example, S. M. Berge, et al., “Pharmaceutical Salts,” J. Phar. Sci., 66: 1-19 (1977)). Moreover, the basic group(s) of the compound of the invention may be reacted with suitable organic or inorganic acids to form salts such as acetate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, hydrobromide, camsylate, carbonate, chloride, clavulanate, citrate, chloride, edetate, edisylate, estolate, esylate, fluoride, fumarate, gluceptate, gluconate, glutamate, glycolylarsanilate, hexylresorcinate, hydrochloride, hydroxynaphthoate, hydroiodide, isothionate, lactate, lactobionate, laurate, maleate, mandelate, mesylate, methylbromide, methylnitrate, methylsulfate, mucate, napsylate, nitrate, oleate, oxalate, palmitate, pantothenate, phosphate, polygalacturonate, salicylate, stearate, subacetate, succinate, tannate, tartrate, tosylate, trifluoroacetate, trifluoromethane sulfonate, and valerate.
Certain compounds of the invention may possess one or more chiral centers, and thus, may exist in optically active forms. Likewise, when the compounds contain an alkenyl or alkenyl group, there exist the possibility of cis- and trans-isomeric forms of the compounds. The R— and S-isomers and mixtures thereof, including racemic mixtures as well as mixtures of cis- and trans-isomers, are contemplated by this invention. Additional asymmetric carbon atoms can be present in a substituent group such as an alkyl group. All such isomers as well as the mixtures thereof are intended to be included in the invention. If a particular stereoisomer is desired, it can be prepared by methods well known in the art by using stereospecific reactions with starting materials which contain the asymmetric centers and are already resolved or, alternatively by methods which lead to mixtures of the stereoisomers and subsequent resolution by known methods. For example, a racemic mixture may be reacted with a single enantiomer of some other compound. This changes the racemic form into a mixture of stereoisomers and diastereomers, because they have different melting points, different boiling points, and different solubilities and can be separated by conventional means, such as crystallization.
Prodrugs are derivatives of the compounds of the invention which have chemically or metabolically cleavable groups and become by solvolysis or under physiological conditions the compounds of the invention which are pharmaceutically active in vivo. Derivatives of the compounds of this invention have activity in both their acid and base derivative forms, but the acid derivative form often offers advantages of solubility, tissue compatibility, or delayed release in a mammalian organism (see, Bundgard, H., Design of Prodrugs, pp. 7-9, 21-24, Elsevier, Amsterdam 1985). Prodrugs include acid derivatives well known to practitioners of the art, such as, for example, esters prepared by reaction of the parent acidic compound with a suitable alcohol, or amides prepared by reaction of the parent acid compound with a suitable amine. Simple aliphatic or aromatic esters derived from acidic groups pendent on the compounds of this invention are preferred prodrugs. In some cases it is desirable to prepare double ester type prodrugs such as (acyloxy) alkyl esters or ((alkoxycarbonyl)oxy)alkyl esters. Particularly preferred esters as prodrugs are methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, tert-butyl, morpholinoethyl, and N,N-diethylglycolamido.
Compounds useful for the invention are prepared following the general procedures outlined below. One of skill in the art is aware that minimal experimentation would be needed to modify the schemes herein provided for the preparation of other compounds of the invention. Scheme 1 provides a general method starting from available dihydroindazole-2-one compound.
According to Scheme 1, 2,3-dihydro-indazole 1 may be reacted with p-nitrophenylchloroformate or other suitable leaving group synthon in the presence of a suitable base to afford the compound 2. The resulting compound 2 is readily reacted with an amine to effect displacement of the p-nitrophenol group affording the carbamoyl compound 3. The carbamoyl compound 3 may be further alkylated or otherwise substituted with a nucleophile at the free ring nitrogen atom to afford an N-substituted dihydroindazole carboxylic acid derivative 4, a compound of the invention.
Alternatively, the dihydro-indazole 1 may be reacted with an isocynate having the desired alkyl, aryl, or other substituent to afford directly the compound 3. The compound 3 is a compound of the invention, which may itself be converted to an N-substituted derivative compound of the formula 4 as discussed above. The isocyanate reagent may be prepared as shown in Scheme 1a below wherein R is represented by the group 5-methylhexyl:
Preparation of the isocyanate may be accomplished by adding a solution of the amine e.g. 5-methylhexylamine, and a proton sponge e.g. triethylamine, dropwise to a cold solution of triphosgene in anhydrous dichloromethane. A detailed procedure for the formation of the isocyanate derivative is provided in the experimental section, or may be found in general organic texts and references therein or is known to one of skill in the art. The resulting isocyanate is typically obtained after acidic workup and purification by chromatography or crystallization.
Benzyl derivatives of compounds of formula I may be prepared as shown in Scheme 2.
The amide component of the compound of formula I is introduced via an isocyanate or amine prepared from the corresponding or nitrile. As shown above, an appropriately substituted aniline is converted to the nitrile 5 following procedures described in the experimental section and/or known to one of skill in the art. For example, the aniline is reacted with tert-butoxynitrite in the presence of copper cyanide in DMSO or other suitable solvent. The initial reaction results in a diazonium intermediate that is displaced by cyanide ion to form the nitrile 5. The nitrile 5 is then reduced to the benzyl amine derivative 6 using for example lithium aluminum hydride in anhydrous ethyl alcohol as a reducing agent. The benzyl amine derivative 6 is then reacted with an activated dihydroindazole 2 having the p-nitrophenoxy leaving group as shown in Scheme 1 and discussed previously.
Alternatively, the amine 6 may be converted to the isocyanate 7 via reaction of the amine with triphosgene in the presence of a proton sponge. The resulting isocyanate 7 is then reacted with an appropriately substituted dihydro-indazole to afford the compound 8, a compound of the invention.
In another procedure, the benzyl amine compound 6 may be reacted directly with a nucleophile source such as compound 2 to afford the desired compound 8 as shown in Scheme 3.
Compounds of formula I wherein the starting material dihydro-indazole is not readily available may be prepared starting with an appropriately substituted 2-aminobenzoic acid as shown in Scheme 4
The reaction to form the dihydro-indazole group starting with an aminobenzoic acid 9 is believed to proceed via a diazonium salt intermediate (Sandmeyer reaction, see March, J., Advanced Organic Chemistry, 3rd Edition, Wiley Interscience, New York, N.Y.). The diazonium intermediate may be formed by adding sodium nitrite to a solution of 9. The diazonium salt intermediate is reduced by stannuous chloride to form a phenylhydrazine intermediate which ring closes in-situ to afford the dihydro-indazole compound 10. Detailed procedures are provided in the experimental section, are known to one of skill in the art, or are readily accessed by one of skill in the art via available literature and reference sources.
The dihydro-indazole compound 10 may then be reacted with isocyanate 7 (from Scheme 3) to afford compound 11, a compound of the invention. The carboxylic acid side chain on the dihydro-indazole 12 may be converted to the methyl ester 13 prior to reaction with an isocyanate derivative to afford-the compound 14. The compound 14 is a compound of the invention.
Protection of side chain substitutents may also be necessary for the purpose of alkylating or substituting on the secondary amine position of the dihydro-indazole as shown in Scheme 5:
As shown in scheme 5, the carboxylic acid side chain of compound 15 may be protected by reaction with diphenylmethyl diazomethane to afford the corresponding ester 16 (diphenylmethylester). Protection of the acid allows for a facile nucleophilic reaction of an alkyl halide (R7X) or other nucleophile source on the free dihydro-indazole ring NH proton of compound 16. The resulting N alkyl or N-substituted compound 17 may be hydrolyzed under acidic conditions to afford the deprotected, N-substituted compound 18.
Compounds of formula I wherein all of R3, R4, R5, and R6 are other than hydrogen are made starting with purchased starting materials having the requisite substituents or by methods known to one of skill in the art.
The dihydro-1H-indazole-5-carboxylic acid derivative compounds described herein are believed to achieve their beneficial therapeutic action principally by direct inhibition of hepatic lipase and/or endothelial lipase activity.
The method of the invention for inhibiting hepatic lipase and/or endothelial lipase activity with a therapeutically effective amount of a dihydro-1H-indazole-5-carboxylic acid derivative compound of Formula (I) as described herein including a combination thereof, a salt or a prodrug or enantiomer derivative thereof.
Another aspect of this invention relates to inhibition or prevention of “Hepatic Lipase-Mediated Diseases” such as hypercholesterolemia, stroke, atherosclerosis and related diseases as described earlier. The method comprises of administering to a mammal (including a human) in need of such treatment a therapeutically effective amount of a dihydro-1H-indazole-5-carboxylic acid derivative compound of the invention.
As previously noted the compounds of this invention are useful for inhibiting hepatic lipase and/or endothelial lipase activity. By the term, “inhibiting” is meant the prevention or therapeutically significant reduction in hepatic lipase and/or endothelial lipase activity by the compounds of the invention. By “pharmaceutically acceptable” it is meant the carrier, diluent or excipient must be compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.
The specific dose of a compound administered according to this invention to obtain therapeutic or ameliorative or prophylactic effect will, of course, be determined by the particular circumstances surrounding the case, including, for example, the compound administered, the route of administration and the condition being treated. Typical daily doses will contain a non-toxic dosage level of from about 0.01 mg/kg to about 50 mg/kg of body weight of an active compound of this invention.
Preferably compounds of the invention per Formula (I) or pharmaceutical formulations containing these compounds are in unit dosage form for administration to a mammal. The unit dosage form can be a capsule or tablet itself, or the appropriate number of any of these. The quantity of Active Ingredient in a unit dose of composition may be varied or adjusted from about 0.1 to about 500 milligrams or more according to the particular treatment involved. It may be appreciated that it may be necessary to make routine variations to the dosage depending on the age and condition of the patient. The dosage will also depend on the route of administration.
The compound of formula I may be administered by a variety of routes including oral, aerosol, transdermal, subcutaneous, intravenous, intramuscular, or intranasal.
Pharmaceutical formulations of the invention are prepared by combining (e.g., mixing) a therapeutically effective amount of the dihydro-1H-indazole-5-carboxylic acid derivative compound of the invention together with a pharmaceutically acceptable carrier or diluent. The present pharmaceutical formulations are prepared by known procedures using well-known and readily available ingredients.
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, or can be in the form of tablets, pills, powders, lozenges, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solid or in a liquid medium), or ointment, containing, for example, up to 10% by weight of the active compound. The compounds of the present invention are preferably formulated prior to administration.
For the pharmaceutical formulations any suitable carrier known in the art can be used. In such a formulation, the carrier may be a solid, liquid, or mixture of a solid and a liquid. For example, for intravenous injection the compounds of the invention may be dissolved in at a concentration of 2 mg/ml in a 4% dextrose/0.5% Na citrate aqueous solution. Solid form formulations include powders, tablets and capsules. A solid carrier can be one or more substance, which may also act as flavoring agents, lubricants, solubilizers, suspending agents, binders, tablet disintegrating agents and encapsulating material.
Tablets for oral administration may contain suitable excipients such as calcium carbonate, sodium carbonate, lactose, calcium phosphate, together with disintegrating agents, such as maize, starch, or alginic acid, and/or binding agents, for example, gelatin or acacia, and lubricating agents such as magnesium stearate, stearic acid, or talc. A preferred tablet formulation for oral administration is one that affords rapid dissolution in the mouth of a patient in need thereof.
In powders the carrier is a finely divided solid, which is in admixture with the finely divided Active Ingredient. In tablets the Active Ingredient is mixed with a carrier having the necessary binding properties in suitable proportions and compacted in the shape and size desired. The powders and tablets preferably contain from about 1 to about 99 weight percent of the Active Ingredient, which is the novel compound of this invention. Suitable solid carriers are magnesium carbonate, magnesium stearate, talc, sugar lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethyl cellulose, low melting waxes, and cocoa butter.
Sterile liquid form formulations include suspensions, emulsions, syrups and elixirs.
The Active Ingredient can be dissolved or suspended in a pharmaceutically acceptable carrier, such as sterile water, sterile organic solvent or a mixture of both. The Active Ingredient can often be dissolved in a suitable organic solvent, for instance aqueous propylene glycol. Other compositions can be made by dispersing the finely divided Active Ingredient in aqueous starch or sodium carboxymethyl cellulose solution or in a suitable oil.
The following pharmaceutical formulations 1 through 8 are illustrative only and are not intended to limit the scope of the invention in any way. “Active Ingredient”, refers to a compound according to Formula (I) or a pharmaceutically acceptable salt, solvate, racemate or enantiomer thereof.
Hard gelatin capsules are prepared using the following ingredients:
A tablet is prepared using the ingredients below:
The components are blended and compressed to form tablets each weighing 665 mg
An aerosol solution is prepared containing the following components:
The active compound is mixed with ethanol and the mixture added to a portion of the propellant 22, cooled to −30° C. and transferred to a filling device. The required amount is then fed to a stainless steel container and diluted with the remainder of the propellant. The valve units are then fitted to the container.
Tablets, each containing 60 mg of Active Ingredient, are made as follows:
The Active Ingredient, starch and cellulose are passed through a No. 45 mesh U.S. sieve and mixed thoroughly. The aqueous solution containing polyvinylpyrrolidone is mixed with the resultant powder, and the mixture then is passed through a No. 14 mesh U.S. sieve. The granules so produced are dried at 50° C. and passed through a No. 18 mesh U.S. sieve. The sodium carboxymethyl starch, magnesium stearate and talc, previously passed through a No. 60 mesh U.S. sieve, are then added to the granules which, after mixing, are compressed on a tablet machine to yield tablets each weighing 150 mg.
Capsules, each containing 80 mg of Active Ingredient, are made as follows:
The Active Ingredient, cellulose, starch, and magnesium stearate are blended, passed through a No. 45 mesh U.S. sieve, and filled into hard gelatin capsules in 200 mg quantities.
Suppositories, each containing 225 mg of Active Ingredient, are made as follows:
The Active Ingredient is passed through a No. 60 mesh U.S. sieve and suspended in the saturated fatty acid glycerides previously melted using the minimum heat necessary. The mixture is then poured into a suppository mold of nominal 2 g capacity and allowed to cool.
Suspensions, each containing 50 mg of Active Ingredient per 5 ml dose, are made as follows:
The Active Ingredient is passed through a No. 45 mesh U.S. sieve and mixed with the sodium carboxymethyl cellulose and syrup to form a smooth paste. The benzoic acid solution, flavor and color are diluted with a portion of the water and added, with stirring. Sufficient water is then added to produce the required volume.
An intravenous formulation may be prepared as follows:
The solution of the above ingredients generally is administered intravenously to a subject at a rate of 1 ml per minute.
The abbreviations, symbols and terms used in the examples have the following meanings.
Ac=acetyl
Anal.=elemental analysis
calcd=calculated
Cpd.=compound
DMF=dimethylformamide
DMSO=dimethylsulfoxide
Et=ethyl
EtOAc=ethyl acetate
EtOH=ethanol
EtSH=ethanethiol
ESIMS=Electrospray Ionization Mass Spectrometry
FAB=Fast Atom Bombardment (Mass Spectroscopy)
FDMS=field desorption mass spectrum
Hex=hexanes
HL=Hepatic Lipase
HPLC=High Performance Liquid Chromatograph
HRMS=high resolution mass spectrum
IR=Infrared Spectrum
Me=methyl
MeI=methyl iodide
MeOH=methanol
MPLC=Medium Pressure Liquid Chromatography
NMR=Nuclear Magnetic Resonance
PPA=polyphosphoric acid
Rochelle's Salt=potassium sodium tartrate
RPHPLC=Reversed Phase High Performance Liquid Chromatography
SiO2=silica gel
SM=starting material
Temp.=temperature
TFA=trifluoroacetic acid
THF=tetrahydrofuran
TLC=thin layer chromatography
Compounds of the present invention were found to be efficacious in-vitro in inhibiting hepatic lipase. Efficacy was determined by testing various compounds of the invention in a hepatic lipase assay discussed below, and disclosed in U.S. patent application Ser. No. 09/609,871 filed Jul. 3, 2000 incorporated herein by reference in its entirety for U.S. Patent office purposes.
Reagents
For Hepes Buffer A, there is 2.4 g Hepes/100 mL water. Therefore 36 grams of Hepes is dissolved in 1500 mL of water. The mix solution's pH is adjusted to pH 8.3 at 37° C. and brought up to 1500 mL with water. 500 mL of Buffer A is retained for the Protein Buffer.
Hepes Buffer B
To the remaining 1000 mL of Hepes Buffer A, 4.49 g of Triton X-100 is added and then the combination mixed on a stir plate. It is optimal that stock Buffer A not be too cold or Triton X-100 will take a long time to go into solution
ThioPEG Substrate Solution
For 0.42 mM substrate stock, use 0.227 mg of thioPEG/mL of Substrate Buffer B. Approximately 20 mg of sn-1 thiol substituted. Phosphatidyl Ethylene Glycol (see Examples for preparation method) is weighed into a vial, such as a scintillation vial. Enough chloroform should be added to make a 2.043 mg/mL solution. Sonicate the solution briefly until well dissolved. Next, pipette 1 mL of chloroform/substrate solution into each scintillation vial. This should give enough substrate for one full 96 well plate. Each vial is dried with nitrogen until solvent removed, swirling each vial simultaneously such that a thin film of substrate will be easily reconstituted in each buffer. Each vial is then frozen.
Daily stock preparation is performed for 9 mL of substrate (one microtiter plate). On the day of the assay, the substrate vial is removed from the freezer and combined with 9 mL of pre-warmed (37° C.) substrate buffer (the final concentration is 0.227 mg/mL). Place the buffer in a 37° C. water bath. Sonicate for 5 minutes or vortex until solution is clear before use.
Enzyme Solution
The enzyme is stored at −80° C. in 100 or 50 μL portions. A 0.406 mg/mL recombinant hepatic lipase stock requires a 50-fold dilution. Therefore, to a 50 μL or 100 μL enzyme aliquot, 2450 μL or 4900 μL, respectively, of substrate Buffer A (protein buffer) should be added. The enzyme should then be stored on ice until ready to use. The protein concentration of enzyme is about 0.406 mg/mL.
DTNB Solution
To make a 20 mg/mL stock solution, 2-3 mg of DTNB is weighed and then mixed with an appropriate amount of 100% DMSO (dimethyl sulfoxide) to make the desired concentration. This mixture is sonicated for five minutes.
The above solution should be diluted 10 fold with substrate Buffer B (concentration now 2 mg/mL). Then to the thioPEG substrate solution, add 60 μl of dilute DTNB per mL of thioPEG substrate solution. Thus for 9 mL of substrate, 540 microliters of dilute DTNB (final concentration in substrate solution=0.11 mg/mL).
Table 1 below shows final assay volumes and concentrations of various components used following the above procedure.
Sample Preparation
The test compound is dissolved in pure DMSO at 1 μM (1000 nM). As shown below in Table 2, assay concentrations are 10, 1, 0.1, 0.33, 0.011, 0.0037, 0.0012 and 0.00041 μM. Table 2 shows the assay concentrations and the corresponding volume of stock and 10% DMSO for each concentration.
Assay Procedure
Using a spectrometer, DTNB is used as a thiol coloring reagent with an incubator temperature of 37° C. Substrate Buffer B is placed in a 37° C. water bath to pre-warm. The substrate is removed from the freezer and 9 mL of substrate Buffer B, 100 mM Hepes, 6.83 mM Tx-100) is added, sonicated for 5 min. and then kept in a 37° C. water bath. Dilutions of the test compound are next made in preparation for assay.
10 μl of the diluted test compound are transferred via pipette into the wells. Control wells receive 10 μl each of 10% DMSO and enzyme solution, while blank wells receive 10 microliters of 10% DMSO and 10 microliters of saline (no enzyme).
Next, DTNB is weighed and diluted to 20 mg/mL with DMSO. The DTNB is then diluted 10 fold with the substrate Buffer B. 540 μl of diluted DTNB is added to 9 ml of ThioPEG and mixed well.
The stock enzyme is diluted with Buffer A. Next, 10 microliters of protein solution is added to each well except the blank, and the wells mixed. The stock solution and test compounds are incubated at 37° C. for 10 min. At 10 minutes, 80 microliters of substrate are added to each well. The plate is then placed in the spectrometer and read at 412 nM every 2 minutes for 30 minutes.
Hepatic lipase (HL) and endothelial lipase (EL) were expressed from AV12 cells. Aliquots from one day's collection of media were stored at −70° C. Activity was measured for both enzymes in conditioned media, (non-purified) where they were tested on the same plate with Thio PEG substrate (0.06 mol fraction, 7.24 mM total lipid), at 37° C. for 30 minutes. The HL, at 1×, had an OD of 14.7. The OD for EL at 1× was 6.029. Therefore, when HL was used in studies where it was compared to EL, the HL was at 0.25× and EL was used at 1×. All experiments were done in triplicate with enzyme from conditioned media.
Kinetic experiments for EL were done varying the total lipid with a constant 0.044 mol fraction determining that a 10 mM total lipid was optimal. In addition, kinetic experiments varying the mol fractions with a constant total lipid showed that 0.03 mol fraction was optimal. Each experiment was run three times.
Experiments to determine proper pH of the substrate to be used with EL were performed at 37° C. with the above-mentioned conditions. The enzyme was tested at pH 7.0, 7.4 and 8.3. The order of addition of reagents/enzyme was as follows: 10 μL of 10% DMSO, 80 μL of substrate and 10 μL of enzyme. Data represents an average of three experiments.
Temperature of the assay was varied from 26.9° C. to 37° C. with the above-mentioned conditions. This was the temperature of the incubation during the 30-minute read. The pH of the substrate was 8.3. The order of addition of reagents/enzyme was as follows: 10 μL of 10% DMSO, 80 μL of substrate and 10 μL of enzyme. Each experiment was run three times. Data is an average of these experiments.
Substrate specificity was determined by testing the activity of HL and EL with Thio Phosphatidylethylene glycol (PEG) and Thio-phosphatidylethanolamine (PE).
Assay conditions of assay were as follows. Both substrates for EL were run at 0.03 mol fraction, 10 mM total lipid. They were dissolved in 100 mM Hepes with 9.95 mM TX100. Both substrates for HL were run at 0.06 mol fraction and 7.25 mM total lipid. They were dissolved in 100 mM Hepes with 6.83 mM Triton X100. The EL enzyme was used at 1× and the HL enzyme was used at 0.25×. The order of addition was as follows: 10 μL of 10% DMSO, 80 μL of substrate and 10 μL of enzyme. The DMSO and substrate were incubated for 10 minutes at 37° C. before the addition of the enzyme. DTNB was added to the substrate prior to addition to the well at 0.096 mg/mL final plate. concentration. The experiments were performed 3 times. Data represents an average of these.
Several compounds were tested with HL and EL to determine their ability to inhibit these enzymes. Conditions of the assay were as mentioned in the previous paragraph except for the order of addition. In these experiments, the order of addition was 10 μL of drug/control, 10 μL of enzyme followed by a 10 minute incubation at 37° C. This was followed by the addition of the 80 μL of Thio PEG substrate containing DTNB. Each experiment was performed 3 times. Data represents an average.
While the present invention has been illustrated above by certain specific embodiments, it is not intended that these specific examples should limit the scope of the invention as described in the appended claims.
All of the products of the Examples described below as well as intermediates used in the following procedures showed satisfactory NMR and IR spectra. They also had the correct mass spectral values.
Propyl isocyanate (88.1 μL, 0.940 mmol) is added dropwise to a stirred suspension of 1,2-dihydro-indazol-3-one (105 mg, 0.783 mmol) in anhydrous THF (3 mL) at ambient temperature under nitrogen. The resultant clear solution turns into a heavy suspension in a minute and the suspension is allowed to stir for 2 hours. After concentration and subsequent chromatography on silica (gradient 0-0.5% CH3OH in CH2Cl2), the title compound 3-1 is obtained as a white solid (156 mg, 91% yield). mp 153.0-155.0° C.; ESIMS m/e-220 (M+H)+. Analysis for C11H13N3O2: calcd: C, 60.26; H, 5.98; N, 19.17; found: C, 60.13; H, 5.91; N, 19.01.
By following similar procedure as described in Example 1, the title compound 3-2 (92% yield) is obtained as a white solid. mp 119.0-120.0° C.; ESIMS m/e 248 (M+H)+. Analysis for C13H17N3O2: calcd: C, 63.14; H, 6.93; N, 16.99; found: C, 62.96; H, 6.84; N, 16.91.
By following similar procedure as described in Example 1, the title compound 3-3 (93% yield) is obtained as a white solid. ESIMS m/e 262 (M+H)+. Analysis for C14H19N3O2: calcd: C, 64.35; H, 7.33; N, 16.08; found: C, 64.06; H, 7.43; N, 15.84.
A solution of 4-nitrophenyl chloroformate (3.17 g, 15.7 mmol) in anhydrous THF (15 mL) is added to a stirred suspension of 3-oxo-1,3-dihydro-indazole (2.01 g, 15.0 mmol) in anhydrous THF (30 mL) at ambient temperature under nitrogen. The resultant mixture is stirred for two hours. Then half of THF is evaporated off at ambient temperature under vacuum. The concentrated suspension is treated with Et2O (25 mL), sonicated and filtered to give the title compound 2 as a white solid (3.26 g, 73% yield). 1H-NMR DMSO-d6) δ 7.18 (t, J=7.8 Hz, 1H), 7.30 (d, J=7.8 Hz, 1H), 7.66 (d, J=8.8 Hz, 2H), 7.67 (t, J=7.8 Hz, 1H), 7.77 (d, J=7.8 Hz, 1H), 8.36 (d, J=8.8 Hz, 2H), 10.87 (s, 1H).
2-Ethylhexyl amine hydrobromide (211 mg, 1.00 mmol) is added to a stirred suspension of 3-oxo-1,3-dihydro-indazole-2-carboxylic acid 4-nitro-phenyl ester 2 (300 mg, 1.00 mmol) in anhydrous THF (10 mL) at ambient temperature under nitrogen. Then triethyl amine (0.140 mL, 1.00 mmol) is added to the mixture and the resultant mixture is allowed to stir for 18 hours. The mixture is concentrated and the crude product is, chromatographed on silica (gradient 0-1% EtOAc in CHCl3) to give the title compound 3-4 (227 mg, 78% yield) as a white solid. ESIMS m/e 290 (M+H)+.
A solution of 5-(methyl)hexylamine (407 mg, 3.53 mmol) and proton sponge (1.51 g, 7.06 mmol) in anhydrous CH2Cl2 (6 mL) is added dropwise to a stirred solution of triphosgene (419 mg, 1.41 mmol) in anhydrous CH2Cl2 (6 mL) at 0° C. The resultant solution is allowed to stir at ambient temperature for 15 minutes. After dilution with CH2Cl2 (40 mL), the mixture is washed with 1N HCl (15×2 mL) and water (15 mL). The organic layer is dried over Na2SO4, filtered and concentrated to give the desired isocyanate 1 (365 mg, 73% yield) as oil. 1H-NMR (CDCl3) δ 0.88 (d, J=6.6 Hz, 6H), 1.16-1.23 (m, 2H), 1.32-1.42 (m, 2H), 1.50-1.62 (m, 3H), 3.29 (t, J=6.6 Hz, 2H).
By following similar procedure as described in Example 1, the title compound 3-5 is obtained as a white solid. mp 134.0-136.0° C.; ESIMS m/e 276 (M+H)+.
By following similar procedure as described in Example 1, the title compound 3-6 (99% yield) is obtained as a white solid. ESIMS m/e 290 (M+H)+. Analysis for C16H23N3O2: calcd: C, 66.41; H, 8.01; N, 14.52; found: C, 66.25; H, 7.90; N, 14.75.
By following similar procedure as described in Example 4, the title compound 3-7 (45% yield) is obtained as a white solid. ESIMS m/e 314 (M+H)+. Analysis for C18H23N3O2: calcd: C, 68.98; H, 7.40; N, 13.41; found: C, 68.99; H, 7.30; N, 13.42.
By following similar procedure as described in Example 4, the title compound 3-8 (71% yield) is obtained as a white solid. ESIMS m/e 274 (M+H)+. Analysis for C15H19N3O2: calcd: C, 65.91; H, 7.01; N, 15.37; found: C, 65.88; H, 6.96; N, 15.05.
Methyl iodide (0.426 mL, 7.15 mmol) is added to a stirred suspension of 3-oxo-1,3-dihydro-indazole-2-carboxylic acid pentylamide 3-2 (70.9 mg, 0.287 mmol) and anhydrous K2CO3 (39.6 mg, 0.287 mmol) in anhydrous THF (3 mL) at ambient temperature under nitrogen. The resultant mixture is stirred for 24 hours. Ethyl acetate (30 mL) and half-saturated aquous NaCl solution (10 mL) are added to the mixture. The organic layer is separated, dried over MgSO4, filtered and concentrated. The crude product is chromatographed on silica (gradient 0-3% CH3OH in CH2Cl2) to give the title compound 4-1 as oil (55.0 mg, 73% yield). ESIMS m/e 262 (M+H)+. Analysis for C14H19N3O2: calcd: C, 64.35; H, 7.33; N, 16.08; found: C, 64.17; H, 7.33; N, 16.04.
By following similar procedure as described in Example 8, the title compound 4-2 is obtained as oil. ESIMS m/e 304 (M+H)+. Analysis for C17H25N3O2: calcd: C, 67.30; H, 8.31; N, 13.85; found: C, 67.30; H, 8.40; N, 13.98.
By following similar procedure as described in Example 8, the title compound 4-3 is obtained as oil (90% yield). ESIMS m/e 338 (M+H)+. Analysis for C20H23N3O2: calcd: C, 71.19; H, 6.87; N, 12.45; found: C, 71.30; H, 6.79; N, 12.59.
By following similar procedure as described in Example 8, the title compound 4-4 is obtained as oil. ESIMS m/e 352 (M+H)+.
By following similar procedure as described in Example 8, the title compound 4-5 is obtained as oil. FDMS m/e 319 (M)+.
Methyl bromoacetate (0.724 mL, 7.65 mmol) is added to a stirred suspension of 3-oxo-1,3-dihydro-indazole-2-carboxylic acid hexylamide 3-3 (200 mg, 0.765 mmol) and anhydrous K2CO3 (159 mg, 1.15 mmol) in anhydrous THF (3 mL) at ambient temperature under nitrogen. The resultant mixture is stirred for 24 hours. Ethyl acetate (30 mL) and half-saturated aqueous NaCl solution (10 mL) are added to the mixture. The organic layer is separated, dried over MgSO4, filtered and concentrated. The crude product is chromatographed on silica (gradient 0-0.25% CH3OH in CH2Cl2) to give the title compound 4-6 as oil (188 mg, 74% yield); ESIMS m/e 334 (M+H)+. Analysis for C17H23N3O4: calcd: C, 61.25; H, 6.95; N, 12.60; found: C, 61.12; H, 6.93; N, 12.64.
By following similar procedure as described in Example 1, the title compound 8-1 (91% yield) is obtained as a white solid. ESIMS m/e 268 (M+H)+. Analysis for C15H13N3O2: calcd: C, 67.41; H, 4.90; N, 15.72; found: C, 67.18; H, 5.05; N. 15.62.
By following similar procedure as described in Example 1, the title compound 8-2 (97% yield) is obtained as a white solid. mp 166.0-168.0° C.; ESIMS m/e 282 (M+H)+. Analysis for C16H15N3O2: calcd: C, 68.31; H, 5.37; N, 14.94; found: C, 67.96; H, 5.22; N, 14.77.
By following similar procedure as described in Example 1, the title compound 8-3 (89% yield) is obtained as a white solid. mp 179.0-180.0° C.; ESIMS m/e 282 (M+H)+. Analysis for C16H15N3O2: calcd: C, 68.31; H, 5.37; N, 14.94; found: C, 68.36; H, 5.48; N, 14.85.
4-Methylbenzyl amine (125 mg, 1.03 mmol) is added dropwise to a stirred suspension of 3-oxo-1,3-dihydro-indazole-2-carboxylic acid 4-nitro-phenyl ester 2 (308 mg, 1.03 mmol) in anhydrous THF (10 mL) at ambient temperature under nitrogen. The resultant mixture is allowed to stir for 18 hours. After concentration, the crude product is chromatographed on silica (gradient 0-0.5% CH3OH in CH2Cl2) to give the title compound 8-4 (210 mg, 72% yield) as a white solid. ESIMS m/e 282 (M+H)+. Analysis for C16H15N3O2: calcd: C, 68.31; H, 5.37; N, 14.94; found: C, 68.04; H, 5.38, N, 14.86.
By following similar procedure as described in Example 18, the title compound 8-5 is obtained as a white solid. ESIMS m/e 336 (M+H)+.
To the ice-cold 2-ethyl-benzonitrile 5-1 (2.01 g, 15.3 mmol) in anhydrous THF (20 mL) is slowly added lithium aluminum hydride (1N in THF, 19.2 mL) under nitrogen. The resultant mixture is allowed to stir at ambient temperature for 16 hr. The reaction mixture is cooled at 0° C. and treated dropwise with MeOH (6 mL), then EtOAc (60 mL) and saturated aqueous Rochelle's salt (60 mL) and water (40 mL) are added to the mixture. The resultant two-layered mixture is stirred vigorously under nitrogen for 1 hour. The organic layer is separated, dried over MgSO4, filtered and concentrated to give the title amine 6-1(2.01 g, crude yield 98%) as light oil. 1H-NMR (CDCl3) δ 1.24 (t, J=7.4 Hz, 3H), 1.54 (br s, 2H), 2.69 (q, J=7.4 Hz, 2H), 3.89 (s, 2H), 7.19-7.24 (m, 3H), 7.31-7.35 (m, 1H); ESIMS m/e 136 (M+H)+.
By following similar procedure as described in Example 18, the title compound 8-6 is obtained as a white solid. ESIMS m/e 294 (M−H)−. Analysis for C17H17N3O2: calcd: C, 69.14; H, 5.80; N, 14.23; found: C, 68.86; H, 5.70; N, 14.25.
By following similar procedure as described in Example 20, the title compound 8-7 is obtained as a white solid. ESIMS m/e 296 (M+H)+. Analysis for C17H17N3O2: calcd: C, 69.14; H, 5.80; N, 14.23; found: C, 68.99; H, 5.82; N, 14.21.
By following similar procedure as described in Example 18, the title compound 8-8 is obtained as a white solid. ESIMS m/e 286 (M+H)+.
By following similar procedure as described in Example 18, the title compound 8-9 is obtained as a white solid. ESIMS m/e 302 (M+H, 35Cl)+. Analysis for C15H12ClN3O2: calcd: C, 59.71; H, 4.01; N, 13.93; found: C, 59.40; H, 3.83; N, 13.75.
By following similar procedure as described in Example 18, the title compound 8-10 is obtained as a white solid. mp 186.0-188.0° C.; ESIMS m/e 298 (M+H)+. Analysis for C16H15N3O3: calcd: C, 64.64; H, 5.09; N. 14.13; found: C, 64.35; H, 4.90; N, 14.13.
By following similar procedure as described in Example 18, the title compound 8-11 is obtained as a white solid. ESIMS m/e 350 (M−H)−. Analysis for C16H12F3N3O3: calcd: C, 54.71; H, 3.44; N, 11.96; found: C, 55.00; H, 3.51; N, 11.88.
By following similar procedure as described in Example 18, the title compound 8-12 is obtained as a white solid. ESIMS m/e 314 (M+H)+. Analysis for C16H15N3O2S: calcd: C, 61.32; H, 4.82; N, 13.41; found: C, 61.02; H, 4.75; N, 13.31.
By following similar procedure as described in Example 18, the title compound 8-13 is obtained as a white solid. ESIMS m/e 281 (M−H)−. Analysis for C15H14N4O2: calcd: C, 63.82; H, 5.00; N, 19.85; found: C, 63.47; H, 5.02; N, 19.58.
By following similar procedure as described in Example 18, the title compound 8-14 is obtained as a white solid. ESIMS m/e 309 (M−H)−. Analysis for C17H18N4O2: calcd: C, 65.79; H, 5.85; N, 18.05; found: C, 65.50; H, 5.85; N, 18.01.
By following similar procedure as described in Example 18, the title compound 8-15 is obtained as a white solid. ESIMS m/e 342 (M−H)−. Analysis for C21H17N3O2; calcd: C, 73.45; H, 4.99; N, 12.24; found: C, 73.07; H, 4.82; N, 12.09.
By following similar procedure as described in Example 18, title compound 8-16 is obtained as a white solid. ESIMS m/e 316 (M−H)−.
By following similar procedure as described in Example 18, the title compound 8-17 is obtained as a white solid. ESIMS m/e 330 (M−H)−. Analysis for C20H17N3O2: calcd: C, 72.49; H, 5.17; N, 12.68; found: C, 72.23; H, 5.10; N, 12.33.
By following similar procedure as described in Example 20, the title compound 8-18 is obtained as a white solid. ESIMS m/e 316 [(M+H)+, 35Cl], 318 [(M+H)+, 37Cl]. Analysis for C16H14ClN3O2: calcd: C, 60.86; H, 4.47; N, 13.31; found: C, 60.73; H, 4.49; N, 13.21.
CuCN (7.80 g, 87.2 mmol) is added to a stirred anhydrous DMSO (70 mL) at 60° C. to form a clear solution, and then followed by the addition of t-BuNO2 (24.0 mL, 202 mmol) all at once. A solution of 2-isopropyl-6-methylaniline (10.0 g, 67.0 mmol) in anhydrous DMSO (30 mL) is added dropwise, via an addition funnel, to the mixture. After the addition is complete, the reaction mixture is allowed to stir for 1 hour. After being cooled to 45° C., the mixture is slowly treated with 5N HCl (100 mL). Five minutes later, the reaction mixture is cooled to ambient temperature before it is extracted with EtOAc/hexane (1:1; 500×2 mL). The combined organic layers are washed with water (100 mL) and brine (100 mL), dried, concentrated in vacuo, then chromatographed on silica (0-5% EtOAc in hexane) to give 8.43 g of the crude nitrile 5-2. IR(CHCl3) 2220 cm−1; 1H-NMR (CDCl3) δ1.30 (d, J=6.9 Hz, 6H), 2.54 (s, 3H), 3.38 (h, J=6.9 Hz, 1H), 7.13 (d, J=7.8 Hz, 1H), 7.20 (d, J=7.8 Hz, 1H), 7.41 (t, J=7.8 Hz, 1H); ESIMS m/e 160 (M+H)+.
To the crude ice-cold nitrile 5-2 (7.74 g, 48.6 mmol) in anhydrous Et2O (70 mL) is slowly added lithium aluminum hydride (1N in Et2O, 97.2 mL) under nitrogen. The resultant mixture is allowed to stir at ambient temperature for 16 hours. Then the reaction mixture is cooled at 0° C. and quenched with MeOH until the gas evolution stops. EtOAc (500 mL) and saturated aqueous Rochelle's salt are added and the two-layered mixture is stirred vigorously under nitrogen for 1 hour to give two relatively clear layers. The organic layer is separated, dried over MgSO4, filtered and concentrated, the crude oil is chromatographed on silica [20% EtOAc in hexane, then 1-2% (4.2 M Me3N in EtOH) in CHCl3]. Amine 6-2 (3.78 g, yield 48%) is obtained as a brown oil. IR(CHCl3) 3300 (br) cm−1; 1H-NMR (CDCl3) δ1.16 (d, J=6.8 Hz, 6H), 1.55 (br s, 2H), 2.33 (s, 3H), 3.28 (h, J=6.8 Hz, 1H), 3.71 (s, 2H), 6.92-6.95 (m, 1H), 7.03-7.10 (m, 2H); ESIMS m/e 164 (M+H)+.
By following similar procedure as described in Example 18, the title compound 8-19 is obtained as a white solid (64% yield). ESIMS m/e 324 (M+H)+. Analysis for C19H21N3O2: calcd: C, 70.57; H, 6.55; N, 12.99; found: C, 70.72; H, 6.55; N, 12.90.
By following similar procedure as described in Example 18, the title compound 8-20 is obtained as a white solid. ESIMS m/e 336 [(M+H)+, 35Cl, 35Cl], 338 [(M+H)30 , 35Cl, 37Cl], 340 [(M+H)+; 37Cl; 37Cl].
By following similar procedure as described in Example 20, the title compound 8-21 is obtained as a white solid. ESIMS m/e 332 [(M+H)+, 35Cl], 334 [(M+H)+, 37Cl]. Analysis for C16H14ClN3O3: calcd: C, 57.93; H, 4.25; N, 12.67; found: C, 58.29; H, 4.05; N, 12.52.
By following similar procedure as described in Example 18, the title compound 8-22 is obtained as a white solid (75% yield). ESIMS m/e 326 (M−H)−. Analysis for C17H17N3O4: calcd: C, 62.38; H, 5.23; N, 12.84; found: C, 62.64; H, 5.20; N, 12.86.
By following similar procedure as described in Example 18, the title compound 8-23 is obtained as a white solid. ESIMS m/e 300 (M+H)+.
By following similar procedure as described in Example 18, the title compound 8-24 is obtained as a white solid (73% yield). ESIMS m/e 326 (M+H)+. Analysis for C17H15N3O4: calcd: C, 62.76; H, 4.65; N, 12.92; found: C, 62.79; H, 4.66; N, 12.86.
By following similar procedure as described in Example 20, the title compound 8-25 is obtained as a white solid. ESIMS m/e 310 (M+H)+. Analysis for C18H19N3O2: calcd: C, 69.88; H, 6.19; N, 13.58; found: C, 69.67; H, 6.13; N, 13.53.
By following similar procedure as described in Example 18, the title compound 8-26 is obtained as a white solid. FDMS m/e 349 (M+, 35Cl, 35Cl), 351 (M+, 35Cl, 37Cl), 353 (M+, 37Cl, 37Cl). Analysis for C16H13Cl2N3O2: calcd: C, 54.88; H, 3.74; N, 12.00; found: C, 54.50; H, 3.75; N, 11.72.
By following similar procedure as described in Example 18, the title compound 8-27 is obtained as a white solid (78% yield). ESIMS m/e 356 (M−H)−. Analysis for C18H19N3O5: calcd: C, 60.50; H, 5.36; N, 11.76; found: C, 60.48; H, 5.35; N, 11.73.
By following similar procedure as described in Example 18, the title compound 8-28 is obtained as a white solid (76% yield). ESIMS m/e 310 (M+H)+. Analysis for C18H19N3O2: calcd: C, 69.88; H, 6.19; N, 13.58; found: C, 69.93; H, 6.11; N, 13.58.
By following similar procedure as described in Example 18, the title compound 8-29 is obtained as a white solid (56% yield). ESIMS m/e 308 (M+H)+.
Concentrated hydrochloric acid (103 mL) is added slowly, in a period of 10 minutes, to a stirred suspension of 2-amino-6-methyl-benzoic acid (5.00 g, 33.1 mmol) in water (56 mL) at −25° C., then a solution of sodium nitrite (2.29 g, 33.1 mmol) in water (28 mL) is added dropwise to the cold suspension. The clear solution is allowed to stir for 40 min at −25° C. before it is added in small portions to a stirred solution of Tin(II) chloride (37.6 g, 198 mmol) in concentrated hydrochloric acid (84 mL) at −35° C. The resultant mixture is stirred for 30 minutes at −30° C., then allowed to warm to ambient temperature where it is stirred for another 16 hours. The mixture is filtered and the white solid is washed with cold THF (200 mL) and vacuum dried to give the title compound 9-1 (2.68 g, yield 55%) as a white solid. FDMS m/e 148 (M)+.
1-Isocyanatomethyl-2-methyl-benzene (0.191 mL, 1.30 mmol) is added dropwise to a stirred suspension of 4-methyl-1,2-dihydro-indazol-3-one 9-1 (200 mg, 1.35 mmol) in anhydrous THF (10 mL) at ambient temperature under nitrogen, and the resultant mixture is allowed to stir for 1 hour. After concentration and subsequent chromatography on silica (gradient 0-0.5% CH3OH in CH2Cl2), the title compound 10-1 (119 mg, 31% yield) is obtained as a white solid. ESIMS m/e 296 (M+H)+.
Concentrated hydrochloric acid (42 mL) is added slowly, in a period of 10 minutes, to a stirred suspension of 2-amino-4-methyl-benzoic acid (2.00 g, 13.2 mmol) in water (25 mL) at −25° C., then a solution of sodium nitrite (0.912 g, 13.2 mmol) in water (11 mL) is added dropwise to the cold suspension. The clear solution is allowed to stir for 40 min at −25° C. before it is added in small portions to a stirred solution of Tin (II) chloride (15.0 g, 79.1 mmol) in concentrated hydrochloric acid (34 mL) at −35° C. The resultant mixture is stirred for 30 minutes at −30° C., then allowed to warm to ambient temperature where it is stirred for another 16 hours. The mixture is filtered and the white solid is washed with cold THF and vacuum dried to give a mixture of 2-hydrazino-4-methyl-benzoic acid and the title compound 9-2 (2.56 g). The white solid (2.48 g) is dissolved in methanol (150 mL) and the solution is heated to reflux under nitrogen for 2 days to give 2.33 g of the title compounds 9-2 (100% crude yield).
By following similar procedure as described in part B of Example 44, the title compound 10-2 is obtained as a white solid (39% yield). mp 235° C. (dec); ESIMS m/e 296 (M+H)+.
Concentrated hydrochloric acid (55 mL) is added slowly, in a period of 10 minutes, to a stirred suspension of 2-amino-6-chloro-benzoic acid (3.00 g, 17.2 mmol) in water (30 mL) at 0° C. The resultant mixture is cooled to −30° C. before it is treated dropwise with a solution of sodium nitrite (1.19 g, 17.2 mmol) in water (15 mL). The clear solution is allowed to stir for 40 min at −25° C. before it is added in small portions to a stirred solution of Tin(II) chloride (19.5 g, 103 mmol) in concentrated hydrochloric acid (44 mL) at −35° C. The resultant mixture is stirred for 30 minutes at −30° C., then allowed to warm to ambient temperature where it is stirred for another 16 hours. The mixture is filtered and the white solid is washed with a small amount of cold THF and vacuum dried to give 2.82 g of the title compounds 9-2 (97% crude yield). ESIMS m/e 169 [(M+H)+, 35Cl], 171 [(M+H)+, 37Cl].
By following similar procedure as described in part B of Example 44, the title compound 10-3 is obtained as a white solid (48% yield). ESIMS m/e 314 [(M−H)−, 35Cl], 316 [(M−H)−, 37Cl].
By following similar procedure as described in part A of Example 45, the title compound 9-4 is obtained as a solid (70% crude yield). 1H-NMR (DMSO-d6) □ 7.25 (d, J=8.3 Hz, 1H), 7.30 (d, J=8.3 Hz, 1H), 7.65 (s, 1H), 8.69 (br s, 2H).
By following similar procedure as described in part B of Example 44, the title compound 10-4 is obtained as a white solid (51% yield). ESIMS m/e 316 [(M+H)+, 35Cl], 318 [(M+H)+, 37Cl]. Analysis for C16H14ClN3O2: calcd: C, 60.86; H, 4.47; N, 13.31; found: C, 60.74; H, 4.46; N, 13.16.
By following similar procedure as described in part A of Example 45, the title compound 9-5 is obtained as a solid (94% crude yield). 1H-NMR (DMSO-d6) δ6.95 (d, J=8.3 Hz, 1H), 7.33 (s, 1H), 7.61 (d, J=8.3 Hz, 1H), 8.50 (br s, 2H).
By following similar procedure as described in part B of Example 44, the title compound 10-5 is obtained as a white solid (44% yield). 1H-NMR (DMSO-d6) □ 2.31 (s, 3H), 4.52 (d, J=5.3 Hz, 2H), 7.11-7.31 (m, 6H), 7.75 (d, J=8.3 Hz, 1H), 8.98 (t, J=5.3 Hz, 1H), 11.62 (s, 1H).
By following similar procedure as described in part A of Example 45, the title compound 9-6 is obtained as a solid. 1H-NMR (DMSO-d6) δ3.87 (s, 3H), 7.16 (t, J=7.8 Hz, 1H), 7.32 (d, J=7.8 Hz, 1H), 7.53 (d, J=7.8 Hz, 1H), 9.03 (s, 1H), 9.74 (br s, 1H).
By following similar procedure as described in part B of Example 44, the title compound 10-6 is obtained as a white solid. ESIMS m/e 312 (M+H)+.
By following similar procedure as described in part A of Example 45, the title compound 9-7 is obtained as a solid (94% crude yield).
By following similar procedure as described in part B of Example 44, the title compound 10-7 is obtained as a white solid. 1H-NMR (DMSO-d6) δ2.31 (s, 3H), 4.51 (d, J=5.9 Hz, 2H), 6.95 (s, 1H), 7.13-7.19 (m, 5H), 7.24-7.27-(m, 1H), 9.07 (t, J=5.9 Hz, 1H), 9.52 (s, 1H), 10.59 (s, 1H).
By following similar procedure as described in part B of Example 44, the title compound 10-8 is obtained as an orange solid (62% yield). ESIMS m/e 325 (M−H)−; Analysis for C16H14N4O{tilde over (4)}.2H2O: calcd: C, 58.25; H, 4.40; N, 16.98; found: C, 58.45; H, 4.27; N, 16.79.
Concentrated hydrochloric acid (220 mL) is added slowly, in a period of 10 minutes, to a stirred suspension of 3-amino-phthalic acid hydrochloride (15.3 g, 70.3 mmol) in water (80 mL) at −10° C. The resultant cold mixture is treated dropwise with a solution of sodium nitrite (4.86 g, 70.3 mmol) in water (60 mL). The clear solution is allowed to stir for 1 hour at −10° C. before it is added in small portions, for a period of 15 minutes, to a stirred solution of Tin(II) chloride (79.8 g, 420 mmol) in concentrated hydrochloric acid (179 mL) at −10° C. The resultant mixture is stirred for 30 minutes at −10° C., then allowed to warm up to ambient temperature where it is stirred for another 16 hours. After filtration and vacuum drying, 11.0 g of the title compounds 11-1 (88% yield) is obtained as a white solid. mp>250° C.; ESIMS m/e 179 (M+H)+; 1H-NMR (DMSO-d6) δ87.49 (dd, J=8.3 and 7.3 Hz, 1H), 7.61 (d, J=8.3 Hz, 1H), 7.70 (d, J=7.3 Hz, 1Hz),
1-Isocyanatomethyl-2-methyl-benzene (0.095 mL, 0.68 mmol) is added dropwise to a stirred suspension of 3-oxo-2,3-dihydro-1H-indazole-4-carboxylic acid 11-1 (121 mg, 0.681 mmol) in anhydrous THF (10 mL) at ambient temperature under nitrogen, and the resultant mixture is allowed to stir for 16 hours. After concentration and subsequent chromatography on silica (0.1% HOAc and 0.5% CH3OH in CHCl3), the title compound 12-1 (85.4 mg, 39% yield) is obtained as a white solid. mp>250° C.; ESIMS m/e 326 (M+H)+. Analysis for C17H15N3O4: calcd: C, 62.76; H, 4.65; N, 12.92; found: C, 62.38; H, 4.35; N, 12.68.
Concentrated hydrochloric acid (30 mL) is added slowly to a stirred suspension of 4-amino-isophthalic acid (0.855 g, 4.71 mmol) in water (45 mL)/THF (5 mL) at −10° C. The resultant cold mixture is treated dropwise with a solution of sodium nitrite (0.325 g, 4.71 mmol) in water (5 mL). The clear solution is allowed to stir for 2 hours at −10° C. before it is added in small portions, for a period of 10 minutes, to a stirred solution of Tin(II) chloride (5.33 g, 28.1 mmol) in concentrated hydrochloric acid (12 mL) at −10° C. The resultant mixture is stirred for 30 minutes at −10° C., then allowed to warm up to ambient temperature where it is stirred for another 1 hour. After filtration and vacuum drying, the white solid is dissolved in DMF (20 mL) and the solution is heated at 120° C. for 2 hours. The mixture is then concentrated to a 5 mL solution at 50° C. under vacuum and diluted with xylene to form a white suspension. After filtration and vacuum drying, 803 mg of the title compounds 11-2 (96% yield) is obtained as a white solid. mp>250° C.; ESIMS m/e 179 (M+H)+; 1H-NMR (DMSO-d6) δ7.31 (d, J=8.8 Hz, 1H), 7.83 (d, J=8.8 Hz, 1H), 8.31 (s, 1H), 8.47 (br s, 1H), 11.92 (br s, 1H).
By following similar procedure as described in part B of Example 52, the title compound 12-2 is obtained as a white solid (25% yield). mp>250° C.; ESIMS m/e 326 (M+H)+.
By following similar procedure as described in Example 53, the title compound 12-3 is obtained as a white solid. mp>250° C.; 1H-NMR (DMSO-d6) δ2.32 (s, 3H), 4.54 (d, J=5.9 Hz, 2H), 7.15-7.19 (m, 3H), 7.27-7.29 (m, 1H), 7.61 (d, J=8.3 Hz, 1H), 7.79 (s, 1H), 7.83 (d, J=8.3 Hz, 1H), 9.06 (t, J=5.9 Hz, 1H), 11.65 (br s, 1H).
By following similar procedure as described in Part A of Example 52, the title compound 11-4 is obtained as a white solid (94% crude yield). mp>250° C.; 1H-NMR (DMSO-d6) δ7.07 (t, J=7.4 Hz, 1H), 7.90 (d, J-7.4 Hz, 2H), 11.58 (br s, 1H).
By following similar procedure as described in Part B of Example 52, the title compound 12-4 is obtained as a white solid (50% yield). mp>250° C.; ESIMS m/e 326 (M+H)+.
A solution of 2-trifluoromethyl-benzylamine (1.00 g, 5.71 mmol) and proton sponge (2.45 g, 11.4 mmol) in anhydrous CH2Cl2 (10 mL) is added dropwise to a stirred solution of triphosgene (678 mg, 2.28 mmol) in anhydrous CH2Cl2 (10 mL) at 0° C. The resultant solution is allowed to stir at ambient temperature for 15 minutes. After dilution with CH2Cl2 (20 mL), the mixture is washed with 1N HCl (15×2 mL) and water (15 mL). The organic layer is dried over Na2SO4, filtered and concentrated to give the desired isocyanate 7-1 (1.07 g, 93% crude yield) as oil. 1H-NMR (CDCl3) δ 4.02 (s, 2H), 7.35 (t, J=7.5 Hz, 1H), 7.52-7.60 (m, 2H), 7.63 (d, J=8.1 Hz, 1H).
A solution of 1-isocyanatomethyl-2-trifluoromethyl-benzene 7-1 (1.07 g, 5.32 mmol) in anhydrous DMF (5 mL) is added to a stirred suspension of 3-oxo-2,3-dihydro-1H-indazole-7-carboxylic acid 11-4 (700 mg, 3.93 mmol) in anhydrous DMF (15 mL) at ambient temperature under nitrogen. The resultant solution is allowed to stir for 2 hours. Then the solution is treated dropwise with 0.5N HCl solution (20 mL) to form a white suspension. After filtration and vacuum drying at 50° C., 855 mg of the title compound 12-5 is obtained as a white solid (57% yield). ESIMS m/e 380 (M+H)+.
By following similar procedure as described in Part A of Example 56, the title compound 7-2 is synthesized from 5-fluoro-2-methyl-benzylamine as oil and used for the subsequent reaction.
A solution of 4-fluoro-2-isocyanatomethyl-1-methyl-benzene 7-2 (313 mg, 1.89 mmol) in anhydrous DMF (2.5 mL) is added to a stirred solution of 3-oxo-2,3-dihydro-1H-indazole-4-carboxylic acid 11-1 (337 mg, 18.9 mmol) in anhydrous DMF (2.5 mL) at ambient temperature under nitrogen. The resultant solution is allowed to stir for 12 hours. Then the solution is concentrated under vacuum and the crude product is treated with CH3OH/CH2Cl2 to from a suspension. The solid is collected by filtration. The solid is then dissolved in DMF and water is added to precipitate out the product. After filtration and vacuum drying at 50° C., the title compound 12-6 is obtained as a white solid. mp 230° C. (dec). ESIMS m/e 344 (M+H)+. Analysis for C17H14FN3O2.0.2H2O: calcd: C, 58.70; H, 4.20; N, 12.08; found: C, 58.42; H, 3.88; N, 12.33.
By following similar procedure as described in Example 57, the title compound 12-7 is obtained as a white solid. mp>250° C.; ESIMS m/e 344 (M+H)+.
By following similar procedure as described in Part A of Example 56, the title compound 7-3 is synthesized from 2,6-dimethyl-benzylamine as oil (83% yield) and used for the subsequent reaction.
By following similar procedure as described in part B of Example 52, the title compound 12-8 is obtained as a solid (27% yield). mp>250° C.; ESIMS m/e 338 (M−H)−.
By following similar procedure as described in part B of Example 52, the title compound 12-9 is obtained as a solid (37% yield). mp>250° C.; ESIMS m/e 340 (M+H)+.
2-Ethyl-6-methyl-benzylamine 6-4 is synthesized by the LiAlH4 reduction of 2-ethyl-6-methyl-benzonitrile according to the procedure described in Part B of Example 33. Then, by following similar procedure as described in Part A of Example 56, the title compound 7-4 is obtained as oil.(91% yield) from compound 6-4. 1H-NMR (DMSO-d6) δ1.16 (t, J=7.6 Hz, 3H), 2.69 (q, J=7.6 Hz, 2H), 4.52 (s, 2H), 7.08 (d, J=7.6 Hz, 2H), 7.16 (t, J=7.6 Hz, 1H).
By following similar procedure as described in part B of Example 52, the title compound 12-10 is obtained as a solid (31% yield). mp>250° C.; ESIMS m/e 354 (M+H)+.
By following similar procedure as described in Example 61, the title compound 12-11 is obtained as a solid. mp>250° C.; ESIMS m/e 354 (M+H)+.
By following similar procedure as described in Example 61, the title compound 12-12 is obtained as a solid. mp>250° C.; ESIMS m/e 354 (M+H)+.
By following similar procedure as described in Example 61, the title compound 12-13 is obtained as a solid (39% yield). mp>250° C.; ESIMS m/e 354 (M+H)+. Analysis for C19H19N3O4: calcd: C; 64.58; H, 5.42; N, 11.89; found: C, 64.36; H, 5.39; N, 11.81.
By following similar procedure as described in Part A of Example 56, the title compound 7-5 is obtained from 2-isopropyl-6-methyl-benzylamine 6-2 as oil (99% yield). 1H-NMR (DMSO-d6) δ1.18 (d, J=6.9 Hz, 6H), 2.35 (s, 3H), 3.24 (h, J=6.9 Hz, 1H), 4.55 (s, 2H), 7.03-7.06 (m, 1H), 7.17-7.23 (m, 2H).
By following similar procedure as described in Part B of Example 52, the title compound 12-14 is obtained as a solid (69% yield). ESIMS m/e 368 (M+H)+.
2-Isocyanatomethyl-1-isopropyl-3-methyl-benzene 7-5 (4.12 g, 21.8 mmol) is added to a stirred suspension of 3-oxo-2,3-dihydro-1H-indazole-7-carboxylic acid 11-4 (2.58 g, 14.5 mmol) in anhydrous DMF (90 mL) at ambient temperature under nitrogen and the resultant clear solution is allowed to stir for 1.5 hours. The solution is treated in small portions with 1N HCl (25 mL) to form a suspension, then followed by the addition of water (100 mL). The white solid is collected by filtration and dried at 50° C. in vacuum oven. A total of 4.69 g of the title compound 12-15 is obtained (88% yield). ESIMS m/e 368 (M+H)+. Analysis for C20H21N3O4: calcd: C, 65.38; H, 5.76; N, 11.44; found: C, 64.99; H, 5.87; N, 11.25.
2-tert-Butyl-6-methyl-benzylamine 6-5 is synthesized from 2-tert-butyl-6-methyl-benzobromide according to the procedure described in Part A and B of Example 33. Then, by following similar procedure as described in Part A of Example 56, the title compound 7-6 is obtained as oil from compound 6-5 (90% yield) and used for the subsequent reaction.
By following similar procedure as described in Example 66, the title compound 12-16 is obtained as solid (85% yield). ESIMS m/e 382 (M+H)+.
4-Butyl-2-methyl-benzylamine 6-6 is synthesized from 4-butyl-2-methyl-benzonitrile according to the procedure described in Part B of Example 33. Then, by following similar procedure as described in Part A of Example 56, the title compound 7-7 is obtained as oil from compound 6-6 (88% yield) and used for the subsequent reaction.
4-Butyl-1-isocyanatomethyl-2-methyl-benzene 7-7 (0.146 mL, 0.842 mmol) is added to a stirred suspension of 3-oxo-2,3-dihydro-1H-indazole-7-carboxylic acid 11-1 (150 mg, 0.842 mmol) in anhydrous DMF (5 mL) at ambient temperature under nitrogen and the resultant mixture is allowed to stir for 16 hours. After concentration the wet solid is suspended in CHCl3 before it is collected by filtration to recover 45 mg of starting 11-1. The filtrate is concentrated and chromatographed on silica (0.1% HOAc and 0.5% CH3OH in CHCl3) to give 36.6 mg of the title compound 12-17 as a yellowish solid (11% yield). mp 173.0-175.0° C.; ESIMS m/e 382 (M+H)+. Analysis for C21H23N3O4: calcd: C, 66.13; H, 6.08; N, 11.02; found: C, 65.87; H, 5.80; N, 10.85.
2,6-Diethyl-benzylamine 6-7 is synthesized from 2-bromo-1,3-diethyl-benzene according to the procedure described in Part A and B of Example 33. Then, by following similar procedure as described in Part A of Example 56, the title compound 7-8 is obtained as oil from compound 6-7 (100% yield). 1H-NMR (DMSO-d6) δ1.16 (t, J=7.8 Hz, 6H), 2.68 (q, J=7.8 Hz, 4H), 4.54 (s, 2H), 7.08 (d, J=7.4 Hz, 2H), 7.21 (t, J=7.4 Hz, 1H).
By following similar procedure as described in Part B of Example 68, the title compound 12-18 is obtained as a white solid (57% yield). mp>250° C.; ESIMS m/e 368 (M+H)+. Analysis for C20H21N3O4: calcd: C, 65.38; H, 5.76; N, 11.44; found: C, 65.18; H, 5.76; N, 11.32.
By following similar procedure as described in Example 69, the title compound 12-19 is obtained as a white solid (42% yield). mp>250° C.; ESIMS m/e 368 (M+H)+. Analysis for C20H21N3O4: calcd: C, 65.38; H, 5.76; N, 11.44; found: C, 65.55, H, 5.86; N, 11.47.
2,6-Diisopropyl-benzylamine 6-8 is synthesized from 2-bromo-1,3-diisopropyl-benzene according to the procedure described in Part A and B of Example 33. Then, by following similar procedure as described in Part A of Example 56, the title compound 7-9 is obtained as oil from compound 6-8 (87% yield). 1H-NMR (DMSO-d6) δ1.18 (d, J=6.8 Hz, 12H), 3.23 (b, J=6.8 Hz, 2H), 4.57 (s, 2H), 7.18 (d, J=7.8 Hz, 2H), 7.28 (t, J=7.8 Hz, 1H).
By following similar procedure as described in Part B of Example 68, the title compound 12-20 is obtained as a yellowish solid (15% yield). ESIMS m/e 396 (M+H)+. Analysis for C22H25N3O4: calcd: C, 66.82; H, 6.37; N, 10:63; found: C, 66.46; H, 6.35; N, 10.60.
By following similar procedure as described in Example 71, the title compound 12-21 is obtained as a white solid (33% yield). mp>250° C.; ESIMS m/e 396 (M+H)+. Analysis for C22H25N3O4: calcd: C, 66.82; H ,6.37; N, 10.63; found: C, 66.51; H, 6.28; N, 10.59.
2,4,6-Trimethyl-benzylamine 6-9 is synthesized from 2-bromo-1,3,5-trimethyl-benzene according to the procedure described in Part A and B of Example 33. Then, by following similar procedure as described in Part A of Example 56, the title compound 7-10 is obtained as oil from compound 6-9 (84% yield). 1H-NMR (CDCl3) δ2.27 (s, 3H), 2.37 (s, 6H), 4.40 (s, 2H), 6.88 (s, 2H).
By following similar procedure as described in Part B of Example 68, the title compound 12-22 is obtained as a white solid (45% yield). ESIMS m/e 354 (M+H)30 .
(4-Methyl-naphthalen-1-yl)-methylamine 6-10 is synthesized from 4-methyl-naphthalene-1-carbonitrile according to the procedure described in Part B of Example 33. Then, by following similar procedure as described in Part A of Example 56, the title compound 7-10 is obtained as oil from compound 6-10 (96% yield). 1H-NMR (DMSO-d6) δ2.64 (s, 3H), 5.00 (s, 2H), 7.35 (d, J=7.0 Hz, 1H), 7.45(d, J=7.0 Hz, 1H), 7.59-7.63 (m, 2H), 8.05-8.08 (m, 2H).
By following similar procedure as described in Part B of Example 68, the title compound 12-23 is obtained as a yellowish solid (24% yield); ESIMS m/e 376 (M+H)+.
By following similar procedure as described in Part A of Example 56, the title compound 7-12 is synthesized from 4-phenyl-butylamine as oil (81% yield) and used for the subsequent reaction.
By following similar procedure as described in Part B of Example 68, the title compound 12-24 is obtained as a white solid (34% yield). mp>250° C.; ESIMS m/e 354 (M+H)+. Analysis for C19H19N3O4: calcd: C, 64.58; H, 5.42; N, 11.89; found: C, 64.42; H, 5.42; N, 11.88.
4-Cyclohexyl-butylamine is synthesized from 4-cyclohexyl-butyronitrile according to the procedure described in Part B of Example 33. Then, by following similar procedure as described in Part A of Example 56, 4-cyclohexyl-butylamine is converted to the title compound 7-13 as oil (91% yield) and used for the subsequent reaction.
By following similar procedure as described in Part 8 of Example 68, the title compound 12-25 is obtained as a white solid. mp 196.0-198.0° C.; ESIMS m/e 360 (M+H)+. Analysis for C19H25N3O40.1H2O: calcd: C, 63.18; H, 7.03; N, 11.63; found: C, 62.96; H, 6.93; N, 11.45:
By following similar procedure as described in Part A of Example 56, the title compound 7-14 is synthesized from 3-phenoxy-propylamine as oil (95% yield) and used for the subsequent reaction.
By following similar procedure as described in Part B of Example 68, the title compound 12-26 is obtained as a white solid. mp 266.0-208.0° C.; ESIMS m/e 356 (M+H)+.
By following similar procedure as described in Part B of Example 52, the title compound 12-27 is obtained as a white solid (40% yield). mp>250° C.; ESIMS m/e 304 (M−H)−.
A solution of diphenyldiazomethane (646 mg, 3.32 mmol) in CH2Cl2 (10 mL) is added dropwise to a stirred suspension of 2-(2-ethyl-6-methyl-benzylcarbamoyl)-3-oxo-2,3-dihydro-1H-indazole-7-carboxylic acid 12-13 (1.12 g, 3.17 mmol) in CH2Cl2 (30 mL) at ambient temperature. The resultant mixture is allowed to stir for 1 hour. The clear solution is concentrated and the crude product is chromatographed on silica (gradient 0-0.5% CH3OH in CHCl3) to give 1.38 g of the title compound 13 (84% yield) as white foam. ESIMS m/e 520 (M+H)+. Analysis for C32H29N3O4: calcd: C, 73.97; H, 5.63; N, 8.09; found: C, 74.28; H, 5.90; N, 7.95.
(Trimethylsilyl)diazomethane (0.42 mL, 2M in hexane) is added dropwise to a stirred solution of 2-(2-ethyl-6-methyl-benzylcarbamoyl)-3-oxo-2,3-dihydro-1H-indazole-7-carboxylic acid benzhydryl ester 13 (424 mg, 0.816 mmol) in CH3OH (5 mL)/CH2Cl2 (10 mL) at ambient temperature. The resultant solution is allowed to stir for 4 hours. The solution is concentrated and the crude product is chromatographed on silica (gradient 1-20% EtOAc in hexane) to give 309 mg of the title compound 14-1 (71% yield) as white foam. ESIMS m/e 534 (M+H)+. Analysis for C33H31N3O4: calcd: C, 74.28; H, 5.86; N, 7.87; found: C, 73.89; H, 5.74; N, 7.65.
Hydrogen chloride gas is bubbled slowly through a stirred solution of 2-(2-ethyl-6-methyl-benzylcarbamoyl)-1-methyl-3-oxo-2,3-dihydro-1H-indazole-7-carboxylic acid benzhydryl ester 14-1 (280 mg, 0.525 mmol) in CH2Cl2 (25 mL) at ambient temperature for a few minutes and the resultant solution is allowed to stir for 1 hour. After concentration, the crude product is chromatographed on silica (gradient 0.1/0.5/99.4 to 0.5/1/98.5 HOAc/CH3OH/CHCl3) to give 135 mg of the title compound 15-1 (70% yield) as a white solid. mp>250° C.; ESIMS m/e 368 (M+H)+.
Ethyl iodide (0.52 mL, 6.5 mmol) and potassium carbonate (44 mg, 0.32 mmol) are added successively to a stirred solution of 2-(2-ethyl-6-methyl-benzylcarbamoyl)-3-oxo-2,3-dihydro-1H-indazole-7-carboxylic acid benzhydryl ester 13 (83.6 mg, 0.161 mmol) in anhydrous DMF (4 mL) at ambient temperature under nitrogen. The resultant mixture is allowed to stir for 3 hours. Xylene (5 mL) is added to the mixture. After filtration and subsequent concentration in vacuo, the crude product is chromatographed on silica (gradient 1-20% EtOAc in hexane) to give 67.8 mg of the title compound 14-2 (77% yield) as white foam. ESIMS m/e 548 (M+H)+.
By following similar procedure as described in Part C of Example 79, the title compound 15-2 is obtained as a white solid (55% yield). mp>250° C.; ESIMS m/e 382 (M+H)+.
By following similar procedure as described in Example 80, the title compound 15-3 is obtained as a white solid (56% yield). mp>250° C.; ESIMS m/e 396 (M+H)+. Analysis for C22H25N3O4.0.2H2O: calcd: C, 66.22; H, 6.42; N, 10.53; found: C, 66.04; H, 6.36; N, 10.45.
tert-Butyl bromoacetate (0.89 mL, 5.8 mmol) and potassium carbonate (0.24 g, 1.7 mmol) are added successively to a stirred solution of 2-(2-ethyl-6-methyl-benzylcarbamoyl)-3-oxo-2,3-dihydro-1H-indazole-7-carboxylic acid benzhydryl ester 13 (0.30 g, 0.58 mmol) in anhydrous DMF (8 mL) at ambient temperature under nitrogen. The resultant mixture is allowed to stir for 1 hour. Xylene (10 mL) is added to the mixture. After filtration and subsequent concentration in vacuo, the crude product is chromatographed on silica (gradient 1-20% EtOAc in hexane) to give 278 mg of the title compound 14-4 (76% yield) as white foam. ESIMS m/e 634 (M+H)+. Analysis for C38H39N3O6.0.2H2O: calcd: C, 71.61; H, 6.23; N, 6.59; found: C, 71.48; H, 6.21; N, 6.62.
By following similar procedure as described in Part C of Example 79, the title compound 15-4 is obtained as a white solid (68%. yield). mp 172.0-174.0° C.; ESIMS m/e 412 (M+H)+. Analysis for C21H21N3O6.0.2H2O: calcd: C, 60.78; H, 5.20; N, 10.12; found: C, 60.51; H, 4.84; N, 9.88.
By following similar procedure as described in Example 82, the title compound 15-5 is obtained as a white solid. mp 220.0-222.0° C.; ESIMS m/e 426 (M+H)+.
By following similar procedure as described in Part A of Example 79, the title compound 16 is obtained as a white solid (88% yield). ESIMS m/e 532 (M−H)−. Analysis for C33H31N3O4.0.2C6H14: calcd: C, 74.57; H, 6.18; N, 7.63; found: C, 74.96; H, 5.78; N, 7.64.
By following similar procedure as described in Part B of Example 79, the title compound 17-1 is obtained as white foam (79% yield). ESIMS m7z/e 548 (M+H)+. Analysis for C34H33N3O4: calcd: C, 74.57; H, 6.07; N, 7.67; found: C, 74.78; H, 6.19; N, 7.65.
By following similar procedure as described in Part C of Example 79, the title compound 18-1 is obtained as a white solid (97% yield). ESIMS m/e 382 (M+H)+.
Ethyl iodide (0.60 mL, 7.5 mmol) and potassium carbonate (104 mg, 0.750 mmol) are added successively to a stirred solution of 2-(2-isopropyl-6-methyl-benzylcarbamoyl)-3-oxo-2,3-dihydro-1H-indazole-7-carboxylic acid benzhydryl ester 16 (200 mg, 0.375 mmol) in anhydrous DMF (4 mL) at ambient temperature under nitrogen. The resultant mixture is allowed to stir for 5 hours. Xylene (5 mL) is added to the mixture. After filtration and subsequent concentration in vacuo, the crude product is chromatographed on silica (gradient 1-15% EtOAc in hexane) to give 167 mg of the title com pound 17-2 (79% yield) as white foam. ESIMS m/e 562 (M+H)+. Analysis for C35H35N3O4: calcd: C, 74.84; H, 6.28; N, 7.48; found: C, 74.60; H, 6.14; N, 7.33.
By following similar procedure as described in Part C of Example 79, the title compound 18-2 is obtained as a white solid (79% yield). ESIMS m/e 396,(M+H)+. Analysis for C22H25N3O4.0.2H2O: calcd: C, 66.22; H, 6.42; N, 10.53; found: C, 66.30; H, 6.36; N, 10.41.
By following similar procedure as described in Part A and B of Example 85, the title compound 18-3 is obtained as a white solid (50% total yield). ESIMS m/e 410 (M+H)+.
By following similar procedure as described in Part A of Example 85, the title compound 17-4 is obtained as white foam (52% total yield). ESIMS m/e 610 [(M+H)+, 35Cl], 612 [(M+H)+, 37Cl].
Triethylsilane (92.2 □L, 0.576 mmol)) and trifluoroacetic acid (0.222 mL, 2.88 mmol) are added successively to a stirred solution of 1-(3-chloro-propyl)-2-(2-isopropyl-6-methyl-benzylcarbamoyl)-3-oxo-2,3-dihydro-1H-indazole-7-carboxylic acid benzhydryl ester 17-4 (176 mg, 0.288 mmol) in anhydrous CH2Cl2 (3 mL) at ambient temperature under nitrogen. The resultant solution is allowed to stir for 2 hours. After concentration, chromatography on silica [gradient 0-2% HOAc in CH3OH (8)/CH2Cl2 (92)] and subsequent crystallization from CH2Cl2/hexane, 126 mg of the title compound 18-4 is obtained as a white solid (99% yield). ESIMS m/e 444 [(M+H)+, 35Cl], 446 [(M+H)+, 37Cl]. Analysis for C23H26ClN3O4: calcd: C, 62.23; H, 5.90; N, 9.47; found: C, 62.01; H, 5.98; N, 9.39.
By following similar procedure as described in Part A and B of Example 87, the title compound 18-5 is obtained as a white solid (86% total yield). ESIMS m/e 406 (M+H)+. Analysis for C23H23N3O4.0.3H2O: calcd: C, 67.24; H, 5.79; N, 10.23; found: C, 67.21; H, 5.67; N, 10.09.
By following similar procedure as described in Part A and B of Example 87, the title compound 18-6 is obtained as a white solid (96% total yield). ESIMS m/e 408 (M+H)+. Analysis for C23H25N3O4.0.6H2O: calcd: C, 67.24; H, 5.79; N, 10.23; found: C, 67.21; H, 5.67; N, 10.09.
By following similar procedure as described in Part A and B of Example 87, the title compound 18-7 is obtained as a white solid (70% total yield). ESIMS m/e 424 (M+H)30 . Analysis for C24H29N3O4: calcd: C, 68.07; H, 6.90; N, 9.92; found: C, 67.85; H, 6.87; N, 9.83.
By following similar procedure as described in Part A and B of Example 87, the title compound 18-8 is obtained as a white solid (77% total yield). ESIMS m/e 407 (M+H)+. Analysis for C22H22N4O4: calcd: C, 65.01; H, 5.46; N, 13.78; found: C, 65.01; H, 5.43; N, 13.71.
By following similar procedure as described in Part A and B of Example 87, the title compound 18-9 is obtained as a white solid (77% total yield). ESIMS m/e 422 (M+H)30 . Analysis for C24H27N3O4.0.2H2O: calcd: C, 67.81; H, 6.50; N, 9.88; found: C, 67.97; H, 6.56; N, 9.58.
By following similar procedure as described in Part A Example 85, the title compound 17-10 is obtained as white foam (61% yield). Analysis for C39H41N3O6.0.1H2O: calcd: C, 72.11; H, 6.39; N, 6.47; found: C, 71.93; H, 6.18; N, 6.27.
By following similar procedure as described in Part C of Example 79, the title compound 18-10 is obtained as a white solid (37% total yield). ESIMS m/e 424 (M−H)−. Analysis for C22H23N3O6.0.4H2O: calcd: C, 61.08; H, 5.54; N, 9.71; found: C, 60.96; H, 5.37; N, 9.60.
By following similar procedure as described in Part A and B of Example 87, the title compound 18-11 is obtained as a white solid. ESIMS m/e 440 (M+H)+. Analysis for C23H25N3O6: calcd: C, 62.86; H, 5.73; N, 9.56; found: C, 62.91; H, 5.78; N, 9.22.
By following similar procedure as described in Part A and B of Example 87, the title compound 18-12 is obtained as a white solid (55% total yield). ESIMS m/e 425 (M+H)+. Analysis for C22H24N4O5.0.35H2O: calcd: C, 61.34; H, 5.78; N, 13.01; found: C, 60.94; H, 5.38; N, 12.84.
By following similar procedure as described in Part A and B of Example 85, the title compound 18-13 is obtained as a white solid (64% total yield). ESIMS m/e 458 (M+H)+. Analysis for C27H27N3O4: calcd: C, 70.88; H, 5.95; N, 9.18; found: C, 70.53; H, 5.93; N,9.10.
By following similar procedure as described in Part A of Example 79, the title compound 19-1 is synthesized from 12-16 as white foam (52% yield). ESIMS m/e 548 (M+H)+, Analysis for C34H33N3O4: calcd: C, 74.57; H, 6.07; N, 7.67; found: C, 74.20; H, 6.05; N, 7.99.
By following similar procedure as described in Part B of Example 79, the title compound 20-1 is synthesized from 19-1 as white foam (81% yield). ESIMS m/e 562 (M+H)+. Analysis for C35H35N3O4: calcd: C, 74.84; H, 6.28; N; 7.48; found: C, 74.78; H, 6.48; N, 7.26.
By following similar procedure as described in Part C of Example 79, the title compound 21-1 is synthesized from 20-1 as a white solid (73% yield). mp>250° C.; ESIMS m/e 396 (M+H)+. Analysis for C22H25N3O4: calcd: C, 66.82; H, 6.37; N, 10.63; found: C, 66.54; H, 6.38; N, 10.54.
Example 98
By following similar procedure as described in Part A, B and C of Example 79, the title compound 21-2 is synthesized from 12-19 as a white solid. mp>250° C.; ESIMS m/e 382 (M+H)+.
Concentrated HCl (10 mL) is added to a stirred suspension of 3-oxo-2,3-dihydro-1H-indazole-4-carboxylic acid 11-1 (5.02 g, 28.2 mmol) in anhydrous CH3OH (50 mL). The resultant mixture is heated to reflux for 4 hours. The mixture is concentrated and the crude product is chromatographed on silica (gradient 3-10% CH3OH in CHCl3) to give 3.01 g of the title compound 22-1 (55% yield) as a white solid. mp>250° C.; ESIMS m/e 193 (M+H)+. Analysis for C9H8N2O3: calcd: C, 56.25; H, 4.20; N, 14.58; found: C, 56.03; H, 4.10; N, 14.41.
By following similar procedure as described in Example 1, the title compound 23-1 is synthesized from compound 22-1 as a white solid (42% yield). mp 63.0-65.0° C.; ESIMS m/e 320 (M+H)+. Analysis for C16H21N3O4.0.2H2O: calcd: C, 59.50; H, 6.68; N, 13.01; found: C, 59.36; H, 6.37; N, 13.16.
Concentrated H2SO4 (1 mL) is added to a stirred suspension of 3-oxo-2,3-dihydro-1H-indazole-4-carboxylic acid 11-1 (6.02 g, 33.8 mmol) in anhydrous allyl alcohol (100 ml). The resultant mixture is heated to reflux for 8 hours. At ambient temperature the mixture is diluted with EtOAc (150 mL) before it is washed with cold water (80 mL×3). The organic layer is dried over MgSO4, filtered and concentrated. After chromatography on silica (gradient 0-5% CH3OH in CHCl3), 4.10 g of the title compound 22-2 (56% yield) is obtained as a yellowish solid. mp 148.0-150.0° C.; ESIMS m/e 219 (M+H)+. Analysis for C11H10N2O3.0.1H2O: calcd: C, 60.05; H, 4.67; N, 12.73; found: C, 60.08; H, 4.45; N, 12.58.
By following similar procedure as described in Example 1, the title compound 23-2 is synthesized from compound 22-2 as a white solid (67% yield). mp 67.0-69.0° C.; ESIMS m/e 346 (M+H)+.
By following similar procedure as described in Part A of Example 100, the title compound 22-3 is synthesized from 3-oxo-2,3-dihydro-1H-indazole-7-carboxylic acid 11-4 as a yellowish solid (48% yield). mp 245.0-246.0° C.; ESIMS m/e 193 (M+H)+.
By following similar procedure as described in Example 1, the title compound 23-3 is synthesized from compound 22-3 as yellowish oil (98% yield). ESIMS m/e 320 (M+H)+. Analysis for C16H21N3O4: calcd: C, 60.18; H, 6.63; N, 13.16; found: C, 60.09; H, 6.57; N, 12.82.
2-Isocyanatomethyl-1-isopropyl-3-methyl-benzene 7-5 (229 mg, 1.21 mmol) is added to a stirred solution of compound 22-1 (250 mg, 1.21 mmol) in anhydrous DMF (5 mL) at ambient temperature under nitrogen. The resultant mixture is allowed to stir for 3 hours. After concentration and subsequent chromatography on silica (gradient 0-1% CH3OH in CH2Cl2), 394 mg of the title compound 24-1 is obtained as a yellowish solid (85% yield). ESIMS m/e 382 (M+H)+. Analysis for C21H23N3O4: calcd: C, 66.13; H, 6.08; N, 11.02; found: C, 66.29; H, 6.20; N, 11.00.
By following similar procedure as described in Example 102, the title compound 24-2 is synthesized from compound 22-3,as a yellowish solid (32% yield). ESIMS m/e 382 (M+H)+. Analysis for C21H23N3O4: calcd: C, 66.13; H, 6.08; N, 11.02; found: C, 66.00; H, 6.21; N, 10.89.
| Filing Document | Filing Date | Country | Kind | 371c Date |
|---|---|---|---|---|
| PCT/US04/06092 | 3/25/2004 | WO | 8/10/2005 |
| Number | Date | Country | |
|---|---|---|---|
| 60459362 | Mar 2003 | US |
