JP2002532525A - Dihydropyrimidine and uses thereof - Google Patents

Abstract

(57) SUMMARY The present invention is directed to dihydropyrimidines, which are selective antagonists of the human α _1A receptor. The invention also relates to the use of these compounds for relaxing the lower urethra tissue, for treating benign prostatic hyperplasia, and for treating any disease in which antagonism of the _α1A receptor may be useful. The present invention further provides a pharmaceutical composition comprising a therapeutically effective amount of the above compound and a pharmaceutically acceptable carrier.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001] This application is related to US Ser. No. 09 / 221,668 filed Dec. 23, 1998 and 1998.
Claim priority of US Provisional Application No. 60 / 113,612, filed December 23, 2012, the contents of which are incorporated herein by reference.

[0002] Throughout this application, various references are cited in parentheses. The disclosures of these publications are hereby incorporated by reference into this application in order to more fully describe the state of the art to which this invention pertains.

[0003]

BACKGROUND OF THE INVENTION

The notation “α _1A ” refers to the cloning subtype previously designated as “α _{1 D} ” as outlined in the receptor and ion channel nomenclature appendix of 1995 (Watson and Girdlestone, 1995). It is a name recently approved by the IUPHAR naming committee. The notation α _1A is used throughout the application and corresponds to the tables and figures to refer to this receptor subtype. At the same time, the receptor, previously designated α _{1 A} , has been _renamed α _1D . This new name is used throughout the present application. Stable cell lines expressing these receptors are described herein, but these cell lines have been deposited with the American Type Culture Collection (ATCC) under the old name (supra). .

[0004] Benign prostatic hyperplasia (BPH), also called benign prostatic hyperplasia, is a progressive condition characterized by nodular thickening of the prostate tissue resulting in urethral disorders. As a result, the frequency of urination increases, nocturnal polyuria occurs, the urine flow is weakened, the start of the urine flow is deteriorated or delayed. Long-term results of BPH can include thickening of bladder smooth muscle, increased incidence of decompensated bladder and urinary tract infections. The specific biochemical, histological and pharmacological properties of prostate adenomas that cause bladder outlet obstruction are not yet known. However, the development of BPH appears to be an inevitable phenomenon for older men. BPH is found in about 70% of men over 70. Currently, in the United States, surgery is the best way to treat BPH (Lepor,
J., Urol. Clinics North Amer., 17, 651 (1990). Over 400,000 prostatectomy cases are performed each year (data since 1986). Clearly, a medicinal alternative to surgery is highly desirable. Limitations of surgery to treat BPH include operative morbidity in older men, persistence or recurrence of disability and irritation, and the significant cost of surgery.

[0005] α-adrenergic receptors (McGrath et al., Med, Res, Rev., 9, 407-533,
(1989) is a specific neuroreceptor protein located in the peripheral and central nervous system of tissues and organs throughout the body. These receptors are important switches for controlling many physiological functions, and therefore represent important targets for drug development. In fact, a number of α-adrenergic drugs have been developed over the last 40 years. Examples include clonidine, phenoxybenzamine and prazosin (treating hypertension), naphazoline (a nasal decongestant) and apraclonidine (treating glaucoma). Alpha-adrenergic agonists are divided into two distinct classes: agonists that mimic the receptor-activating properties of the endogenous neurotransmitter norepinephrine (clonidine and naphazoline are agonists), and block by acting on norepinephrine Antagonists (phenoxybenzamine and prazosin are antagonists). Many of these drugs are effective but also have undesirable side effects (eg, clonidine has an antihypertensive effect,
Causing dryness and sedation of the mouth).

[0006] During the past 15 years, an increase in scientific research has led to a more accurate understanding of α-adrenergic receptors and their drugs. 1977
Years before, it was known that one α-adrenergic receptor was present. Between 1977 and 1988, the central and peripheral nervous system
That kind of α- adrenergic receptor (α ₁ and α ₂₎ is present was observed in the Scientific Committee. Since 1988, novel techniques in molecular biology have enabled at least six α-adrenergic receptors in the central and peripheral nervous system: α _1A (new name), α _1B , α _1D (new name) , Α _2A , α _2B
And alpha _2C were identified (Bylund, DB, FASEB J., 6, 832 (1992)). In many cases, it is not exactly known which physiological responses of the body are controlled by each of these receptors. Furthermore, current α-adrenergic drugs are not at all selective for certain α-adrenergic receptors. Many of these drugs have deleterious side effects, which may be due to their poor selectivity for α-adrenergic receptors.

[0007] Since the mid 1970's, non-selective α-antagonists have been prescribed to treat BPH. In 1976, M. Caine et al. (Brit. J. Urol., 48, 255 (1976))
Reported that phenoxybenzamine, a non-selective α-antagonist, was useful in alleviating the symptoms of BPH. This agent can produce its effect by interacting with α-receptors located in the prostate. However, the drug also produces considerable side effects such as dizziness and asthenia, which severely limits its use in treating patients on a chronic basis. More recently, the α-adrenergic antagonists prazosin and terazosin have also been found to be useful in treating BPH. However, these drugs also have deleterious side effects. Recently, alpha _1A receptor has been discovered that participate in mediating contraction of human prostate smooth muscle (Gluchowski, C. et al., WO94 / 10989, 1994: Forray , C. et al., Mol
Phrmacol. 45, 703, 1994). This finding indicates that α _1A antagonists may be effective drugs with fewer side effects for the treatment of BPH. Further studies indicate that _α1A receptors may also be present in other lower urinary tract tissues, such as urethral smooth muscle (
Ford et al., Br. J. Pharmacol., 114, 24P, (1995)).

[0008] The present invention is directed to dihydropyrimidine compounds that are selective antagonists of the cloned human α _1A receptor. The invention also relates to the use of these compounds for treating benign prostatic hyperplasia and for treating any disease for which antagonism of the α _1A receptor may be useful.

[0009]

Summary of the Invention

 The invention has the following structure:

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Wherein R ₁ is —OCH ₃ or OH; R ₂ is —CH ₂ OH, —CH ₂ OCH ₃ or —COOH; and R ₁ and R ₂ are both a 5-membered lactone ring. R ₃ is — (CH ₂ ) ₃ OH,

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Is selected from the group consisting of [0011] (wherein, R ₃ is when R ₁ is OH

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Or a pharmaceutically acceptable salt thereof.

The present invention also provides the following structure:

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And a compound having the formula:

The present invention provides a pharmaceutical composition comprising a therapeutically effective amount of any one of the compounds described herein and a pharmaceutically acceptable carrier.

The present invention also provides a subject suffering from benign prostatic hyperplasia comprising administering to the patient an amount of any one of the compounds described herein effective to treat benign prostatic hyperplasia. A method of treating is provided.

The present invention provides that lower urinary tract tissue comprises contacting the lower urinary tract tissue with an amount of any one of the compounds described herein effective to relax the lower urinary tract tissue. To provide a way to relax.

The present invention inhibits contraction of prostate tissue in a patient comprising administering an amount of any one of the compounds described herein effective to inhibit contraction of prostate tissue. Provide a way.

The invention is susceptible to treatment by antagonism of the α _1A receptor, comprising administering to a patient an amount of any one of the compounds described herein effective to treat the disease. To provide a method for treating certain diseases.

The present invention provides a pharmaceutical composition comprising a therapeutically effective amount of any one of the compounds described herein in combination with a therapeutically effective amount of finasteride and a pharmaceutically acceptable carrier. provide.

The present invention provides a method of making a pharmaceutical composition comprising combining a therapeutically effective amount of any one of the compounds described herein with a pharmaceutically acceptable carrier.

[0022]

DETAILED DESCRIPTION OF THE INVENTION

 The invention has the following structure:

Embedded image

Wherein R ₁ is —OCH ₃ or OH; R ₂ is —CH ₂ OH, —CH ₂ OCH ₃ or —COOH; and R ₁ and R ₂ are both a 5-membered lactone ring. R ₃ is — (CH ₂ ) ₃ OH,

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(Where R ₁ is OH 2, R ₃ is selected from the group consisting of

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Or a pharmaceutically acceptable salt thereof.

The present invention also provides (+) and (−) enantiomers of all compounds described herein.

The present invention further provides the cis and trans isomers of all compounds described herein. Note that "cis" and "trans" correspond to the relevant stereochemistry, as determined, for example, by NOE (nuclear Overhauser effect) experiments.

The compounds of the present invention may exist as enantiomers, diastereomers, isomers,
That is, two or more compounds may be present in a racemic mixture. Further, the compounds of the present invention are preferably at least 80% pure, more preferably 90% pure, and more preferably 95% pure.

In one embodiment, the compound has the following structure:

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[0030]

In another embodiment of the present invention, the compound is

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## EQU1 ##

In a further embodiment of the present invention, the compound is

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[0034]

In a still further embodiment of the invention, the compound is

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Is as follows.

In an embodiment of the present invention, the compound is

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Is as follows.

In an embodiment of the present invention, R ₃ is

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Selected from the group consisting of:

In a further embodiment of the present invention, the compound has the following structure:

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Has the following.

In an embodiment of the present invention, the compound is

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Is as follows.

In yet another embodiment of the present invention, the compound is

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Is as follows.

In yet another embodiment of the present invention, the compound is

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Is as follows.

The present invention also provides the following structure:

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There is provided a compound having the formula:

The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of any of the compounds described herein and a pharmaceutically acceptable carrier. In the context of the present invention, a "therapeutically effective amount" is any amount of a compound that, when administered to a patient suffering from a disease for which the compound is effective, causes reduction, alleviation, or amelioration of the disease.

In one embodiment, the therapeutically effective amount is from about 0.01 mg per patient per day to about 500 mg per patient per day, preferably about 0.5 mg per patient per day.
The amount is from 1 mg to about 60 mg per patient per day, more preferably from about 1 mg per patient per day to about 30 mg per patient per day.

In the practice of the present invention, a “pharmaceutically acceptable carrier” is any physiological carrier known to those skilled in the art that is useful in formulating a pharmaceutical composition.

[0054] In one embodiment, the pharmaceutical carrier will be liquid and the pharmaceutical composition will be in the form of a solution. In another embodiment, the pharmaceutically acceptable carrier is a solid and the composition is in tablet or powder form. In a further embodiment, the pharmaceutical carrier is a gel and the composition is in the form of a suppository or cream. In still further embodiments, the compounds may be formulated as part of a pharmaceutically acceptable transdermal patch.

The solid carrier includes a flavoring agent, a lubricant, a solubilizer, a suspending agent, a bulking agent, a lubricant,
It can include one or more substances that can also act as tabletting aids, binders or tablet disintegrants, which can also be encapsulating materials. In powders, the carrier is a finely divided solid which is in a mixture with the finely divided active component. In tablets, the active component is mixed with the carrier having the necessary compression properties in suitable proportions and compacted in the shape and size desired. The powders and tablets preferably contain up to 99% of the active ingredient. Suitable solid carriers include, for example, calcium phosphate, magnesium stearate, talc, sugar, lactose, dextrin, starch, gelatin, cellulose, polyvinylpyrrolidine, low melting waxes and ion exchange resins.

[0056] Liquid carriers are used in preparing solutions, suspensions, emulsions, syrups, elixirs and pressurized compositions. The active ingredient may be dissolved or suspended in water, an organic solvent, a mixture of both, or a pharmaceutically acceptable liquid carrier such as a pharmaceutically acceptable oil or fat. Liquid carriers include solubilizers, emulsifiers, buffers, preservatives, sweeteners,
Other suitable pharmaceutical additives such as flavoring agents, suspending agents, thickeners, coloring agents, viscosity modifiers, stabilizers or osmotic pressure regulators may be included. Suitable examples of liquid carriers for oral and parenteral administration include water (partially including the abovementioned additives, for example, cellulose derivatives, preferably sodium carboxymethyl cellulose solution) alcohols (monohydric and polyhydric alcohols) And their derivatives, and oils such as coconut and peanut oils. Also, for parenteral administration, the carrier may be an oily ester such as ethyl oleate and isopropyl myristate. Sterile liquid carriers are useful in sterile liquid form compositions for parenteral administration. The liquid carrier for pressurized compositions can be halogenated hydrocarbon or other pharmaceutically acceptable propellant.

Liquid pharmaceutical compositions that are sterile solutions or suspensions can be administered, for example, intramuscularly, intrathecally,
Available for epidural, intraperitoneal or subcutaneous injection. Sterile solutions may also be administered intravenously. The compounds may be prepared as sterile solid compositions that can be dissolved or suspended at the time of administration using sterile water, saline or other suitable sterile injectable media. Carriers are intended to include necessary and inert binders, suspending agents, lubricants, flavoring agents, sweeteners, preservatives, dyes and coatings.

The compound may comprise other solutes or suspending agents, such as saline or glucose, bile salts, gum arabic, gelatin, sufficient to produce an isotonic solution.
It may be administered orally in the form of a sterile solution or suspension containing sorbitan monooleate, polysorbate 80 (oleate ester of sorbitol and its anhydride copolymerized with ethylene oxide), and the like.

The compounds may be administered orally in either liquid or solid composition form. Compositions suitable for oral administration include solid forms such as pills, capsules, granules, tablets and powders and liquid forms such as solutions, syrups, elixirs and suspensions. Useful forms for parenteral administration include sterile solutions, emulsions and suspensions.

The optimal dose to be administered may be determined by one skilled in the art and will depend on the particular compound employed, the strength of the formulation, the mode of administration and the progression of the disease condition. The age of the patient,
Additional factors depending on the particular patient being treated, including weight, gender, diet and time of administration, will require dosage adjustment.

In a further embodiment of the present invention, any one of the compounds described herein
Species do not further cause a decrease in blood pressure at doses effective to reduce prostatic hypertrophy.

The present invention treats a patient suffering from benign prostatic hyperplasia (BPH), comprising administering to the patient any one of the compounds described herein effective to treat BPH. Provide a way. The present invention further provides compounds that do not further cause a decrease in blood pressure at doses effective to reduce BPH. In one preferred embodiment, the compound achieves treatment of BPH by relaxing lower urinary tract tissue, particularly where lower urinary tract tissue is urethral smooth muscle.

In the practice of the present invention, “lower urinary tract tissue” is used to include the prostatic capsule, prostatic urethra, bladder neck, urethral smooth muscle and prostate smooth muscle.

The invention also comprises administering to a patient one of the compounds described herein in combination with a 5-α-reductase inhibitor effective to treat benign prostatic hyperplasia. A method for treating a patient suffering from hypertrophy is provided. In one embodiment,
The 5-α-reductase inhibitor is finasteride.

The present invention relates to relaxing lower urinary tract tissue, comprising contacting lower urinary tract tissue with an amount of one of the compounds described herein effective to relax lower urinary tract tissue. A further method is provided. In one embodiment, the lower urinary tract tissue is urethral smooth muscle. In one preferred embodiment, the compound does not further cause a decrease in blood pressure when effective to relax lower urinary tract tissue.

The present invention provides a method of inhibiting prostate tissue contraction comprising administering to a patient an amount of any of the compounds described herein effective to inhibit prostate tissue contraction. Provide further. In one preferred embodiment, the prostate tissue is prostate smooth muscle and the compound does not further cause a decrease in blood pressure.

The present invention also provides one of the compounds described herein that is effective in treating a disease.
There is provided a method of treating a disease susceptible to treatment by antagonism of the _α1A receptor, comprising administering a species to a patient.

The present invention provides a pharmaceutical composition comprising a therapeutically effective amount of any one of the compounds described herein in combination with a therapeutically effective amount of finasteride and a pharmaceutically acceptable carrier. Offer things. The invention also provides a pharmaceutical composition comprising any one of the compounds described herein present in an amount from about 0.01 mg to about 500 mg, and wherein the therapeutically effective amount of finasteride is about 5 mg. In one embodiment, the therapeutically effective amount of the compound is from about 0.1 mg to about 60 mg, and the therapeutically effective amount of finasteride is about 5 mg. In another embodiment,
A therapeutically effective amount of a compound is an amount from about 1 mg to about 30 mg, and a therapeutically effective amount of finasteride is about 5 mg.

The present invention provides a pharmaceutical composition prepared by combining a therapeutically effective amount of any of the compounds described herein, and a pharmaceutically acceptable carrier.

The present invention also provides a pharmaceutical composition prepared by combining a therapeutically effective amount of any of the compounds described herein with a therapeutically acceptable amount of finasteride and a pharmaceutically acceptable carrier. Offer things.

Included in the present invention are pharmaceutically acceptable salts and complexes of all compounds described herein. The salts include, but are not limited to, the following acids and bases: inorganic acids, including hydrochloric acid, hydrofluoric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, and boric acid; Including trifluoroacetic acid, formic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, maleic acid, citric acid, methanesulfonic acid, trifluoromethanesulfonic acid, benzoic acid, glycolic acid, glycolic acid, lactic acid, and mandelic acid Organic acids; inorganic bases including ammonia and hydrazine; and organic bases including methylamine, ethylamine, hydroxyethylamine, propylamine, dimethylamine, diethylamine, trimethylamine, triethylamine, ethylenediamine, hydroethylamine, morpholine, piperazine and guanidine Is mentioned. The present invention further provides hydrates and polymorphs of all compounds described herein.

The present invention includes within its scope prodrugs of the compounds of this invention. In general, such prodrugs will be functional derivatives of the compounds of the present invention which are readily convertible in vivo into the desired compound. Accordingly, in the treatment method of the present invention, the term “administration” refers to a specific compound in vivo after administration to a patient with the specifically disclosed compound or not specifically disclosed, for the various symptoms described. Should include treatment with compounds that convert to Routine methods for the selection and preparation of suitable prodrug derivatives are described, for example, in Design of Prodrugs, H. Bundgaard Elsevier, eds., 1985.

The present invention further includes metabolites of the compounds of the present invention. Metabolites include active species that form upon introduction of a compound of the present invention into a biological environment.

It will be readily apparent to one of skill in the art that appropriate biological assays may be used to determine the therapeutic potential of the claimed compounds for treating the above-mentioned disorders.

The present invention will be better understood from the experimental details below. However, one of ordinary skill in the art will readily appreciate that the specific methods and results discussed are merely illustrative of the invention as more fully described in the claims that follow.

[0076]

[Experiment Details]

I. Synthesis of compounds 1-9 A. Synthesis of compound 1 (+)-1,2,3,6-tetrahydro-1- (3-hydroxypropyl) carboxamide-
5-methoxycarbonyl-4-methoxymethyl-6- (3,4-difluorophenyl) -2-
Oxo-pyrimidine a) 5-methoxycarbonyl-4-methoxymethyl-1,2,3,6-tetrahydro-2-oxo-6- (3,4-difluorophenyl) -pyrimidine methyl 4- (300 mL) Methoxyacetoacetate (50 g, 0.351 mol),
3,4-difluorobenzaldehyde (51.39 g, 0.351 mmol), and urea (31
.64 g (0.527 mol) to a well stirred mixture of copper (I) oxide (5.06 g, 0.035 mol) and acetic acid (2.05 mL) at room temperature, followed by boron trifluoride diethyl etherate (56 mL, 0.456 mol). ) Was added dropwise. The mixture was stirred and refluxed for 8 hours, whereupon TLC (1/1 EtOAc / hexane) indicated that the reaction was complete. It was cooled, poured into a mixture of ice and sodium bicarbonate (100 g), and the resulting mixture was filtered through celite. The celite pad was washed with dichloromethane (400mL). The organic layer was separated from the filtrate and the aqueous layer was extracted with additional dichloromethane (3 × 300 mL). The combined organic extracts were dried (sodium sulfate) and the solvent was evaporated.
The crude product was purified by flash column on silica gel using 50% ethyl acetate in hexane, then ethyl acetate as eluent to give the product as pale yellow foam,
This was triturated with hexane to give a white powder (103.3 g, 94%). ¹ H NMR (CDCl ₃ ) δ3.476 (s, 3H), 3.651 (s, 3H), 4.653 (s, 2H), 5.39 (s, 1H), 6.
60 (br s, 1H, NH), 7.00-7.20 (m, 3H), 7.72 (br s, 1H, NH).

B) (+)-5-methoxycarbonyl-4-methoxymethyl-1,2,3,6-tetrahydro-2
-Oxo-6- (3,4-difluorophenyl) -pyrimidine Intermediate in racemic 5-methoxycarbonyl-4-methoxymethyl-1,2,3,6-tetrahydro-2-oxo-6- (3,4-difluoro Phenyl) -pyrimidine is purified by chiral HPLC
[Chiralcel OD 20 x 250 mm # 369-703-30604; l 254 nm; hexane / ethanol
90/10; 85 mg per injection; retention time of the desired enantiomer: 16.94 min, first peak of the enantiomer eluting], separated by (+)-5-methoxycarbonyl-4-methoxymethyl-1, 2,3,6-tetrahydro-2-oxo-6- (3,4-difluorophenyl) -pyrimidine (40-42% by weight of the desired enantiomer isolated from the racemate); [α] _D =
+83.8 (C = 0.5, chloroform).

C) (+)-5-Methoxycarbonyl-4-methoxymethyl-1,2,3,6-tetrahydro-2
-Oxo-6- (3,4-difluorophenyl) -1-[(4-nitrophenyloxy) carbonyl] pyrimidine (+)-5-methoxycarbonyl- in anhydrous THF (20 mL) at -78 ° C under argon atmosphere Four
-Methoxymethyl-1,2,3,6-tetrahydro-2-oxo-6- (3,4-difluorophenyl) -pyrimidine (1.98 g, 6.34 mmol) solution of lithium hexamethyldisilazide in THF The solution (1M, 18 mL, 18 mmol) was added over 2-3 minutes and the mixture was stirred for 10 minutes. This solution was added to a stirred solution of 4-nitrophenyl chloroformate (4.47 g, 22.2 mmol) in THF (20 mL) at -78 ° C via cannula over 6 minutes. Stirring was continued for 10 minutes, the mixture was poured onto ice (50 g) and extracted with chloroform (2 × 50 mL). The combined extracts were dried (sodium sulfate) and the solvent was evaporated. Residue in hexane / ethyl acetate (4: 1 to 3.5: 1)
Was purified by flash column chromatography using as eluent. The product was obtained as a yellow syrup, which was triturated with hexane to give a white powder (2.4 g, 79%). _{^{1 H NMR (CDCl 3) δ3.522}} (s, 3H), 3.744 (s,
3H), 4.65-4.80 (q, J = 16.5 Hz, 2H), 6.323 (s, 1H), 7.10-7.30 (m, 4H), 7.358 (
d, J = 9 Hz, 2H), 8.273 (d, J = 9 Hz, 2H).

D) (+)-1,2,3,6-tetrahydro-1- (3-hydroxypropyl) carboxamido-5-methoxycarbonyl-4-methoxymethyl-6- (3,4-difluorophenyl) -2
-Oxo-pyrimidine (+)-1,2,3,6-tetrahydro-6- (3,4-difluorophenyl) -5-methoxycarbonyl-4-methoxymethyl-1- [N- in dichloromethane (5 mL) To a solution of (4-nitrophenoxy) carbonyl] -2-oxopyrimidine (100 mg, 0.209 mmol) was added 3-aminopropanol (0.2 mL, 2.62 mmol) at room temperature. The resulting mixture was stirred for 2 hours and the crude product was purified by column chromatography on silica gel using EtOAc as eluent (82 mg, 95%). ¹ H-NMR (300 MHZ, CDCl ₃ )
δ1.67-1.75 (m, 2H), 3.41-3.49 (m, 2H), 3.48 (s, 3H), 3.55-3.62 (m, 2H), 3.7
1 (s, 3H), 4.68 (s, 2H), 6.67 (s, 1H), 7.06-7.20 (m, 3H), 7.76 (s, 1H), 8.92 (
t, J = 7 Hz, 1H).

B. Synthesis of Compound 2: (+)-1,2,3,6-tetrahydro-1- {N- [4- (2-pyridyl) piperidin-1-yl]
Propyl｝ carboxamide-5-methoxycarbonyl-4-methoxymethyl-6- (3,
4-difluorophenyl) -2-oxopyrimidine-pyridine-N-oxidehydrochloride a) 4'-t-butoxycarbonyl-1 ', 2', 3 ', 4', 5 ', 6'-hexahydro [2, 4 '] Bipyridinyl 1', 2 ', 3', 4 ', 5', 6'-Hexahydrochloride in dichloromethane (100mL)
To the [2,4 '] bipyridinyl (1.7 g, 10 mmol) solution was added di-t-butyl dicarbonate (2.2 g, 10 mmol) followed by a saturated aqueous sodium bicarbonate solution (100 mL). The mixture was stirred vigorously for 2 days, and when the layers separated, the aqueous layer was diluted with dichloromethane (1
× 100 mL). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo to give crude 4'-t-butoxycarbonyl-1 ', 2', 3 ', 4', 5 ', 6'-
Hexahydro [2,4 '] bipyridinyl was obtained and used as in step b.

B) 4′-t-butoxycarbonyl-2-oxo-1 ′, 2 ′, 3 ′, 4 ′, 5 ′, 6
'-Hexahydro [2,4'] bipyridinyl 4 '-t-butoxycarbonyl-1', 2 ', 3', 4 in dichloromethane (60 mL)
', 5', 6 '-Hexahydro [2,4'] bipyridinyl (10 mmol)
Cooled to ° C. and treated with 3-chloroperoxybenzoic acid (4.3 g, 12.5 mmol). The solution was warmed to ambient temperature and then stirred for 18 hours. The reaction was then treated with saturated aqueous sodium bicarbonate and the layers were separated. The organic layer was dried over magnesium sulfate, filtered, and concentrated in vacuo. The crude material was purified by pressure silica gel chromatography (1-3% methanol in dichloromethane) to give 4'-t-butoxycarbonyl-2-oxo-1 ', 2', 3 ', 4', 5 ', 6 '-Hexahydro [2,4'] bipyridinyl was obtained. FABMS (M + H) = 279.

C) 2-oxo-1 ′, 2 ′, 3 ′, 4 ′, 5 ′, 6′-hexahydro [2,4 ′] bipyridinyl 4′-t-butoxycarbonyl- in ethyl acetate (150 mL) 2-oxo-1 ', 2',
A solution of 3 ', 4', 5 ', 6'-hexahydro [2,4'] bipyridinyl (2.1 g, 7.6 mmol) was cooled to -78 ° C. Hydrogen chloride gas was bubbled through the solution for 20 minutes, then the reaction was allowed to warm to ambient temperature. The separated solid was collected by filtration and dried in vacuo to give the product as the hydrochloride salt. The free base was isolated by standard methods to give 2-oxo-1 ', 2', 3 ', 4', 5 ', 6'-hexahydro [2,4'] bipyridinyl.

D) Nt-butoxycarbonyl-3- (3 ′, 4 ′, 5 ′, 6′-tetrahydro-2-oxo-2′H- [2,4 ′] bipyridinyl-1′-yl) propylamine 2-oxo-1 ', 2', 3 ', 4', 5 ', 6'-hexahydro [2
, 4 ']-Bipyridinyl (0.86g, 4.8mmol) solution in 3-bromopropyl-t-
Butoxycarbonylamine (1.4 g, 5.8 mmol) and cesium carbonate (1.2 g,
3.6 mmol) was added under argon. The solution was warmed to 50 ° C. and stirred for 2 days, then the reaction was cooled to ambient temperature and volatiles were removed in vacuo. The residue was taken up in dichloromethane (100 mL), washed with saturated sodium carbonate (1 × 50 mL), dried over magnesium sulfate, filtered and concentrated in vacuo. 1-3% methanol crude material in dichloromethane containing pressurized silica gel chromatography (0.5% NH ₄ OH
) To give Nt-butoxycarbonyl-3- (3 ', 4', 5 ', 6'-tetrahydro-2-oxo-2'H- [2,4'] bipyridinyl-1'-yl) propylamine I got FABMS (M + H) = 336.

E) 3- (3 ′, 4 ′, 5 ′, 6′-tetrahydro-2-oxo-2′H- [2,4 ′] bipyridinyl-1′-yl) propylamine dihydrochloride Ethyl acetate Nt-butoxycarbonyl-3- (3 ', 4', 5 ', 6'-
Tetrahydro-2-oxo-2′H- [2,4 ′] bipyridinyl-1′-yl) propylamine (0.63 g, 1.9 mmol) was cooled to −78 ° C. After bubbling hydrogen chloride gas through the solution for 10 minutes, the reaction was warmed to ambient temperature. The separated solid was collected by filtration and dried in vacuo to give the product.

F) (+)-1,2,3,6-tetrahydro-1- {N- [4- (2-pyridyl) piperidin-1-yl] propyl} carboxamido-5-methoxycarbonyl-4- Methoxymethyl-6
-(3,4-Difluorophenyl) -2-oxopyrimidine-pyridine-N-oxide hydrochloride (+)-5-Methoxycarbonyl-4-methoxymethyl-1,2,3,6-in THF (20 mL) Tetrahydro-2-oxo-6- (3,4-difluorophenyl) -1-[(4-nitrophenyloxy) carbonyl] pyrimidine (200 mg, 0.42 mmol) (as described in IA (c)) and 3- ( 3 ', 4', 5 ', 6' -tetrahydro-2-oxo-2 'H- [2,4'
Bipyridinyl-1′-yl) propylamine dihydrochloride (100 mg, 0.42) was stirred at room temperature for 24 hours. The solvent was evaporated and the residue was purified by flash chromatography on silica gel (dichloromethane: MeOH: 2M ammonia in MeOH, 980: 10: 10 to 940: 30: 30) to give the desired product (252 mg, 98%). MH + 574.

C. Synthesis of Compound 3: 4- (2-pyridyl) -1- {3- [5-carboxymethyl-4-methoxymethyl-2-oxo
-6 (S)-(3,4-difluorophenyl) -1,2,3,6-tetrahydropyrimidin-1-yl]
Carboxyaminopropyl} pyridinium chloride a) 1- (3-Aminopropyl) -4- (2-pyridyl) pyridinium bromide hydrobromide bromide hydrobromide 2,4'dipyridyl (820 g, 5.25 mol) in DMF (5.0 L) and 3-Bromopropylamine hydrogen bromide (1400 g, 6.39 mol) was heated to 95 ° C. for 8 hours. The reaction mixture was cooled to room temperature and methyl t-butyl ether (3.7 L) was added over 3 hours. The slurry was stirred for 1 hour and filtered. The solid was washed with MTBE / DMF (1: 1, 4.2 L) and dried to give 1- (3-aminopropyl) -4- (2-pyridyl) pyridinium bromide hydrobromide as a brown solid.

B) 4- (2-pyridyl) -1- {3- [5-carboxymethyl-4-methoxymethyl-2-
Oxo-6 (S)-(3,4-difluorophenyl) -1,2,3,6-tetrahydropyrimidine-1-
Yl] carboxyaminopropyl} pyridinium chloride 1- (3-aminopropyl) -4- (2-pyridyl) pyridinium bromide hydrobromide (227 mg, 0.60 mmol), (+)-5-methoxycarbonyl-4-methoxymethyl-1 , 2,3,6-Tetrahydro-2-oxo-6- (3,4-difluorophenyl) -1-[(4-nitrophenyloxy) carbonyl] pyrimidine (287 mg, 0.60 mmol) (I.
A. (c)), and triethylamine (350 mL, 2.5 mmol) in T
Combined in 8 mL of HF and 8 mL of DMF. The yellow mixture was stirred at ambient temperature for 1 hour and then concentrated in vacuo. The resulting oil was purified using a reverse-phase C18 cartridge column and a 0.
Water containing 1% TFA: Waters Delta Prep System using acetonitrile gradient
And purified. The product fractions were evaporated to dryness and the residue obtained was treated with HCl / ethyl acetate. The mixture was evaporated to dryness and the residue was reconcentrated from ethyl ether, then triturated with ethyl ether and filtered to give the title compound. Mp: 60 to 85 ° C. (foam) NMR: Consistent with structure HPLC: purity 99% FAB MS: M + H @ m / e = 552.2 _{Analysis: C 28 H 28 F 2 N} 5 O 5 C1 · HCl · 0.80 H 2 Calculated for O: C, 52.64; H, 4.83
N, 10.96. Found: C, 52.67; H, 4.68; N, 10.77.

D. Synthesis of Compound 4 (+)-4- (2-pyridyl) -1- {3- [5-carboxymethyl-4-methoxymethyl-2-oxo-6- (3,4-difluorophenyl ) -1,2,3,6-tetrahydropyrimidin-1-ylcarbonylamino] propyl} piperidine-1-oxide a) 1- (3-aminopropyl) -4- (2-pyridyl) pyridinium bromide hydrobromide DMF ( 2,4′-Dipyridyl (25 g, 160 mmol) and 3-bromopropylamine hydrobromide (35 g, 160 mmol) in 60 mL) were heated at 90-95 ° C. for 10 minutes. After cooling to room temperature, anhydrous ether (500 mL) was added to the mixture and the white solid formed was filtered, washed with Et ₂ O and dried to give 60 g (100%). ¹ H NMR (3
00 MHZ, DMSO-d ₆ ) δ2.35-2.44 (m, 2H), 3.08-343 (nu, 2H), 4.76-4.81 (m, 2H),
7.58 (dd, J = 4.8 Hz, J = 7.5 Hz, 1H), 8.03 (dt, 1.8 Hz, J = 7.8 Hz, 1H), 8.32 (
d, 3 7.8 Hz, 1H) , 8.77-8.81 (m, 3H), 9.12 (d, J = 6.3 Hz, 2H) _{Analysis:. C 13 H 16 N 3} Br.HBr.0.5H Calcd for ₂ O: C, 40.65; H, 4.72 ;. N, 10.94. Found: C
, 40.83 H, 4.37; N, 11.05.

B) 3- (3 ′, 6′-dihydro-2′-H- [2,4 ′] bipyridinyl-1′-yl) propylamine 1- (3-aminopropyl) in MeOH (150 mL) -4- (2-Pyridyl) pyridinium bromide hydrobromide (6 g, 16 mmol) NaBH ₄ (2 g, 53 mmol)
Was added portionwise over 2 hours at 0-5 ° C. The reaction mixture was stirred overnight at room temperature and the solvent was evaporated. The residue was suspended in ether (200 mL) and 50% NaOH solution (100 mL
). The ether layer was separated and the aqueous layer was extracted with more ether (2 × 50 mL). The combined ether extracts were dried over potassium carbonate, the solvent was removed and 3- (3 ′, 6′-dihydro-2′-H- [2,4 ′] bipyridinyl-1′-yl was obtained as an oil.
) Propylamine was obtained (3.48g). This was used directly in the next step without purification.

C) 3-Aminopropyl-4- (2-pyridyl) piperidine 3- (3 ′, 6′-dihydro-2′-H- [2,4 ′] bipyridinyl-1 in MeOH (40 mL)
To a solution of '-yl) propylamine (unpurified 3.48 g, 15.9 mmol) was added 1.0 g of Perlman's catalyst. After hydrogenating the suspension at 120 psi for 10 hours, the reaction mixture was filtered through a pad of celite to remove the solvent. The residue was silica gel (30 g) using CH ₂ Cl ₂ in MeOH / methanol / 2M NH ₃ (90: 8: 4 to 90:40:40) as eluent
Purification by column chromatography on a column. The product was obtained as a pale yellow oil (3.21 g, 91%). _{^{1 H NMR δ (CD 3 OD}} ) 1.50-1.99 (m, 10H), 2.02-2.06 (m,
2H), 2.37-2.75 (m, 3H), 3.02-3.06 (br in, 2H), 7.05-7.09 (m, 4H), 7.16 (dt,
J = 0.9 Hz, J = 8.7 Hz, 1H), 8.48 (dd, J = 0.9 Hz, J = 4.2 Hz, 1H).

D) (+)-6- (3,4-difluorophenyl) -1- {N- [4- (2-pyridyl) piperidine-1-
Yl] propyl]} carboxamido-5-methoxycarbonyl-4-methoxymethyl-2-oxo 1,2,3,6-tetrahydropyrimidine (+)-5-methoxycarbonyl-4-methoxymethyl in THF (20 mL) -1,2,3,6-tetrahydro-2-oxo-6- (3,4-difluorophenyl) -1-[(4-nitrophenyloxy) carbonyl] pyrimidine (as described in IA (c)) ( 2.38 g, 5 mmol
)) And 3-aminopropyl-4- (2-pyridyl) piperidine (1.206 g, 5.5 mmol) were stirred at room temperature for 12 hours. Evaporate the solvent and elute the residue with ethyl acetate (100 mL)
Was dissolved again. This was washed with ice-cold 1N NaOH (4 × 50 mL), brine (2 × 50 mL) and dried over potassium carbonate. The solvent was evaporated under reduced pressure and the residue was purified by flash chromatography on silica gel (dichloromethane: MeOH: 2M ammonia in MeOH, 980: 10: 10 to 940: 30: 30) to give 2.45 g of very pure product
(88%) and a slightly impure fraction 0.30 g (10%) were obtained as foam. ¹ H NMR (CDCl ₃ )
δ1.60-2.00 (m, 6H), 2.05-2.15 (m, 2H), 2.38-2.43 (br t, 2H), 2.65-2.80 (m,
1H), 3.05-3.06 (br d, 2H), 3.30-3.45 (m, 2H), 3.477 (s, 3H), 3.704 (s, 3H),
4.678 (s, 2H), 6.68 (s, 1H), 7.05-7.20 (m, 5H), 7.58-7.63 (dt, 1H), 7.702 (s,
1H, NH), 8.50-8.52 (dd, 1H), 8.875 (brt, 1H).

(E) (+)-4- (2-pyridyl) -1- {3- [5-carboxymethyl-4-methoxymethyl-2-
Oxo-6- (3,4-difluorophenyl) -1,2,3,6-tetrahydropyrimidine-1-
Ylcarbonylamino] propyl} piperidine-1-oxide (+)-6 (S)-(3,4-difluorophenyl) -1- {3- [4- (2-pyridyl) piperidine-1- in 4 mL of methanol Yl] propylamino} carbonyl-5-methoxycarbonyl-4-methoxymethyl-2-oxo-1,2,3,6-tetrahydropyrimidine (205 mg, 0
.37 mmol) to the solution was added 50% hydrogen peroxide in water (75 mL, 1.30 mmol), and 1
Stirred for 0 days. The reaction was quenched by the addition of platinum black until the evolution of oxygen ceased. The reaction slurry was filtered through celite and the filtrate was evaporated to dryness to give a pale yellow oil. The resulting oil was purified by "flash" chromatography (methylene chloride: methanol: ammonium hydroxide 94: 6: 0.6). Product fractions were evaporated to dryness and re-concentrated several times from ethyl ether to give the title compound as a yellow foam. Mp: 89 to 115 ° C. (foam) NMR: Consistent with structure HPLC: purity 100% FAB MS: M + H @ m / e = 574 _{Analysis: C 28 H 33 F 2 N} 5 O 6 · 0.60H 2 O · 0.10 Calculated for Et ₂ O: C, 57.63; H, 6.0
0; N, 11.83. Found: C, 57.64; H, 5.83; N, 11.80.

E. Synthesis of Compound 5 (+)-1,2,3,6-tetrahydro-1- {N- [4- (2-pyridyl) -4-hydroxypiperidin-1-yl] propyl} carboxamide -5-Methoxycarbonyl-4-methoxymethyl-6- (3,4-difluorophenyl) -2-oxopyrimidine dihydrochloride a) N-benzyl-4-hydroxy-4- (2-pyridyl) piperidine THF (120 mL) To a stirred solution of 2-bromopyridine (10 mmol) in 2.5 g of a 2.5 M solution of n-butyllithium in hexane (4.5 mL, 11.25 mmol) at -78 ° C under argon was injected over 5 minutes. After 25 minutes, 1-benzylpiperidone (10 mmol)
Was added as is. After 30 minutes, the mixture was quenched by addition of aqueous NH ₄ Cl (50 mL). The mixture was diluted with ethyl acetate (200 mL), washed with brine (150 mL), dried (Na ₂ SO _4). The solvent was evaporated and the residue was purified by column chromatography (silica gel, EtOAc / hexane 20% -60%) to give the desired product (1.7
8g, 66.41%). ¹ H-NMR was consistent with the structure.

B) 4-Hydroxy-4- (2-pyridyl) piperidine N-benzyl-4-hydroxy-4- (2-pyridyl) piperidine (1.2 g, 4.48 mmol), 20% Pd-C (0.2 g) And a mixture of EtOH (100 mL) was stirred at H ₂ (150 psi) for 36 h. The catalyst was filtered off and washed with more ethanol. The solvent was evaporated to give pure 4-hydroxy-4- (2-pyridyl) piperidine (0.75 g, 94%).

C) 3- [4-Hydroxy-4- (2-pyridyl) piperidin-1-yl] propylamine 4-Hydroxy-4- (2-pyridyl) piperidine in acetone (100 mL) and THF (20 mL) (0.730 g, 4.1 mmol), K ₂ CO ₃ (0.8 g) and KI (0.1 g) were added to N-bromopropylphthalimide (4.5 mmol) and the mixture was stirred for 14 hours. The crude material was purified by column chromatography (silica gel, MeOH in _{CH 2 Cl 2: MeOH: 2M} NH 3 90: 4: 1 to 75: 0: 25) product obtained as purified by,
A phthalimide protected side chain resulted (1.376 g, 92%). To this was added a solution of concentrated hydrochloric acid and water (2: 1, 25 mL) and the mixture was heated for 20 hours. The reaction mixture was then washed and extracted with ether (4 × 50 mL) to remove the formed phthalic acid. The water was evaporated from the aqueous layer to give the desired product as the hydrochloride salt (1.10 g, 100%). ¹ H-NM
R was consistent with this product. This intermediate was treated with 6N NaOH and converted to the free base before use in the next step.

D) (+)-1,2,3,6-tetrahydro-1- {N- [4- (2-pyridyl) -4-hydroxypiperidin-1-yl] propyl} carboxamido-5-methoxy Carbonyl-4-methoxymethyl-6- (3,4-difluorophenyl) -2-oxopyrimidinedihydrochloride (+)-5-Methoxycarbonyl-4-methoxymethyl-1 in dichloromethane (5 mL)
, 2,3,6-tetrahydro-2-oxo-6- (3,4-difluorophenyl) -1-[(4-nitrophenyloxy) carbonyl] pyrimidine (as described in IA (c)) (201 mg, 0
.421 mmol) solution, a solution of 3- [4-hydroxy-4- (2-pyridyl) piperidin-1-yl] propylamine (160 mg, 0.729 mmol) was added at room temperature. The obtained mixture was stirred for 2 hours, and then concentrated to obtain a residue. The residue was purified by column chromatography (chloroform / MeOH / 2N ammonia in MeOH = 100: 12: 6) to give the desired product (212 mg, 88%). [Α] _D = + 109 ° C = 0.245 in CHCl ₃
). This compound was converted to the hydrochloride salt by dissolving in 1N HCl in ether and then concentrated to dryness. 135-138 ° C. _{Analysis: C 29 H 33 F 2 N} 5 O 6 -2HCl (0.4CH 2 Cl 2 + 1.0H 2 O) Calcd for:. C, 47.61; H, 5.60; N, 9.77 Found: C, .48.39; H, 5.45; N,
^{9.76. 1 H-NMR (300} MHZ, CDCl 3) δ1.57-1.68 (m, 2H), 1.75-1.82 (m, 2H), 2.02
-2.16 (m, 2H), 2.45-2.55 (m, 4H), 2.82-2.86 (m, 2H), 3.35-3.45 (m, 2H), 3.46
(s, 3H), 3.68 (s, 3H), 4.67 (s, 25 2H), 6.67 (s, 1H), 7.02-7.20 (m, 3H), 7.3
8 (d, J = 8 Hz, 1H), 7.66-7.79 (m, 2H), 8.48-8.50 (d, J = 5 Hz, 1H), 8.92 (t, J =
7 Hz, 1H).

F. Synthesis of Compound 6 (+)-2-oxo-3- {N- [4- (2-pyridyl) piperidin-1-yl] propyl} carboxamido-4- (3,4-difluorophenyl ) -5-Methoxycarbonyl-6-carboxy-1,2,3,6-tetrahydropyrimidine a) Trimethylsilylethyl oxalate (13.64 g, 151.4 mmol), trimethylsilylethanol (43.4 ml
, 303 mmol) and 0.5 ml of concentrated sulfuric acid in 300 ml of benzene.
Heated to reflux using a Dean-Stark apparatus. After 18 hours, the reaction mixture was cooled and diluted with 200 ml of ethyl acetate. The reaction mixture was washed with 1N aqueous sodium hydroxide (2 × 100 ml), water (1 × 100 ml) and brine. Dried
(Magnesium sulfate) The organic phase was rotary evaporated to yield 9 g of crude product as an oil. Chromatography on silica gel (chloroform elution) 5.
61 g was obtained.

B) Methyltrimethylsilylethyl oxaloacetate A solution of 1.78 ml of methyl acetate (22.38 mmol) in 25 ml of tetrahydrofuran is cooled to -78 ° C. under nitrogen and lithium bis (trimethylsilyl) amide 24
Treated with .6 ml (24.62 mmol). The reaction mixture was stirred for 1 hour and cannulated into a solution of trimethylsilylethyl oxalate in 25 ml of tetrahydrofuran at -78 ° C. After 3.5 hours, the reaction mixture was quenched with water and ethyl acetate 15
Diluted with 0 ml. The organic phase was separated and washed with 1N hydrochloric acid and brine. Concentration of the dried (magnesium sulfate) extract gave an oil which was used in the next step without further purification.

C) Methyl 2-[(3,4-difluorophenyl) methylene] -trimethylsilylethyl oxaloacetate Methyltrimethylsilylethyl oxaloacetate (4.13 g, 16.78 mmol)
), 3,4-difluorobenzaldehyde (1.85 ml, 16.78 mmol), piperidine (66 ml, 0.67 mmol), and acetic acid (201 ml, 3.52 mmol) were mixed in 50 ml of benzene at ambient temperature. The reaction mixture was then added to Dean-Stark (Dean-S
Reflux in the apparatus for 2.5 hours. The reaction mixture was cooled, diluted with 100 ml of ethyl acetate and washed with water and brine. The combined organic extracts were dried over magnesium sulfate and concentrated to give 7 g of a mobile yellow liquid which was used in the next step without further purification.

D) 2-Methoxy-4- (3,4-difluorophenyl) -5-methoxycarbonyl-6-trimethylsilylethylcarbonyl-1,6-dihydropyrimidine Crude methyl 2 in 50 ml of anhydrous N, N-dimethylformamide 3.7 g (10 mmol) of-[(3,4-difluorophenyl) methylene] -trimethylsilylethyl oxaloacetate, O-methylisourea hemisulfate (2.23 g, 13 mmol), and sodium bicarbonate (3.27 g, 39 mmol) ) Was stirred at 55 ° C. for 6 hours. The reaction mixture was cooled, filtered and concentrated in vacuo. The residue was dissolved in ethyl acetate, and the resulting solution was washed with water. Dry the organic phase (sodium sulfate) and concentrate to about 3 g in the crude foam
The title compound was obtained. This material was used in the next step without further purification.

E) 4- (3,4-difluorophenyl) -5-methoxycarbonyl-6-trimethylsilylethylcarbonyl-1,2,3,6-tetrahydropyrimidin-2-one 2-methoxy-4 in crude foam -(3,4-difluorophenyl) -5-methoxycarbonyl
-6-Trimethylsilylethylcarbonyl-1,6-dihydropyrimidine (3 g, 7 mmol) was dissolved in 20 ml of tetrahydrofuran and treated with 2 ml of 12N hydrochloric acid. The resulting reaction mixture was stirred at ambient temperature for 2.5 hours. The pH of the reaction mixture was adjusted to 7 with 2N lithium hydroxide solution and the reaction mixture was concentrated in vacuo to leave an aqueous phase. The aqueous layer was extracted with ethyl acetate. The combined organic extracts were washed with brine, dried (magnesium sulfate) and concentrated. The crude product was purified using preparative centrifugation thin layer chromatography (hexane-ethyl acetate elution) to give 900 mg of the title compound. ¹ H-NMR (C
DCl ₃ ) d 0.06 (s, 9H), 1.07 (m, 2H), 3.63 (s, 3H), 4.36 (m, 2H), 5.38 (d, 1H),
5.73 (br. S, 1H), 7.02-7.2 (m, 3H), 7.52 (br. S, 1H).

F) 2-oxo-3-[(4-nitrophenyloxy) carbonyl] -4- (3,4-difluorophenyl) -5-methoxycarbonyl-6-trimethylsilylethylcarbonyl-1,
2,3,6-tetrahydropyrimidine 4- (3,4-difluorophenyl) -5-methoxycarbonyl-6-trimethylsilylethylcarbonyl-1,2,3,6-tetrahydropyrimidin-2-one (25 ml of anhydrous tetrahydrofuran) (478 mg, 1.16 mmol) To the solution was added 0.7 ml (1.39 mmol) of lithium diisopropylamide at -78 ° C under nitrogen. After the addition was complete, the reaction mixture was stirred for 1 hour and cannulated into a solution of 280 mg (1.39 mmol) of 4-nitrophenylchloroformate in 15 ml of tetrahydrofuran. The reaction was stirred for an additional hour, warmed to room temperature, and partitioned between 100 ml of ethyl acetate and 50 ml of a 1N sodium carbonate solution. The organic phase was washed once more with sodium carbonate and then with brine. The combined organic extracts were dried (sodium sulfate) and concentrated to give a yellow semi-solid. The crude product was flash chromatographed on silica gel (chloroform-acetone elution, 9: 1, v / v) to give the title compound as a pale yellow solid.

G) (±) -2-oxo-3- {N- [4- (2-pyridyl) piperidin-1-yl] propyl}
Carboxamido-4- (3,4-difluorophenyl) -5-methoxycarbonyl-5-
Trimethylsilylethylcarbonyl-1,2,3,6-tetrahydropyrimidine 3-aminopropyl-4- (2-pyridyl) piperidine (240 mg, 1.1 mmol) and 2-oxo-3-[(4-nitrophenyloxy) carbonyl ] -4- (3,4-Difluorophenyl) -5-methoxycarbonyl-6-trimethylsilylethylcarbonyl-1,2,
3,6-Tetrahydropyrimidine (577 mg, 1.1 mmol) was combined at ambient temperature in 10 ml of anhydrous tetrahydrofuran. The resulting solution was stirred for 2 hours and then diluted with 50 ml of ethyl acetate. The reaction mixture was then washed sequentially with 1N hydrochloric acid, 10% sodium carbonate solution, 1N hydrochloric acid, and brine. The combined organic extracts were dried (sodium sulfate) and concentrated to give 550 mg of crude product. This material was purified by preparative centrifugation thin layer chromatography on silica gel (chloroform-methanol elution) to give 228 mg of the title compound. ¹ H NMR (CDCl ₃ ) d 0.06 (s, 9H), 1.07 (m, 2H)
, 1.78 (m, 2H), 1.91 (m, 4H), 2.1 (m, 2H), 2.45 (m, 2H), 2.71 (rn 1H), 3.05 (m
, 2H), 3.31 (ddd, 1H), 3.42. (Ddd, 1), 3.72 (s, -3H) 4.32 (m, 2H), 6.60 (s, 1
H) 7.05-7.35 (m, 6H), 7.61 (ddd, 1H), 851 (d, 1H), 887 (dd, 1H).

H) (+)-2-oxo-3- {N- [4- (2-pyridyl) piperidin-1-yl] propyl}
Carboxamido-4- (3,4-difluorophenyl) -5-methoxycarbonyl-6-trimethylsilylethylcarbonyl-1,2,3,6-tetrahydropyrimidine Racemic 2-oxo-3- {N- [4- (2-pyridyl) piperidin-1-yl] propyl} carboxamido-4- (3,4-difluorophenyl) -5-methoxycarbonyl-6-trimethylsilylethylcarbonyl-1,2,3,6-tetrahydropyrimidine HPLC (Chiralpak AD, 250 x 20 mm; l 315 nm; isocratic 80% hexane: 20%
2-propanol (containing 0.2% diethylamine); flow rate = 6.0 ml / min]. Recover the first of the two eluting enantiomers to yield the title compound.
110 mg were obtained. Analytical chiral HPLC (Chiralpak AD, 250 x 4.6 mm; l 280 nm; isocratic 80% hexane-20% 2-propanol (containing 0.1% diethylamine); flow rate = 2.
0 ml / min] indicated that the title compound was 98.66% pure.

I) (+)-2-oxo-3- {N- [4- (2-pyridyl) piperidin-1-yl] propyl}
Carboxamido-4- (3,4-difluorophenyl) -5-methoxycarbonyl-6-carboxy-1,2,3,6-tetrahydropyrimidine (+)-2-oxo-3- {N- [4- ( 2-pyridyl) piperidin-1-yl] propyl} carboxamido-4- (3,4-difluorophenyl) -5-methoxycarbonyl-6-trimethylsilylethylcarbonyl-1,2,3,6-tetrahydropyrimidine (110 mg, 0.1
(6 mmol) in 2 ml of anhydrous tetrahydrofuran.
Treated with 33 mL (0.33 mmol) of 1 M tetrabutylammonium fluoride solution. The reaction mixture was protected from moisture and heated to 60 ° C. for 2 hours. All volatiles are removed under reduced pressure and the remaining material is separated by preparative thin-layer chromatography (ethyl acetate-
Purification by pyridine-acetic acid-water eluting with 90: 10: 1: 1, v / v) gave the title compound as an off-white solid. HPLC: 95.5%. ¹ H-NMR (DMSO-d ₆ ) d 1.7-1.9 (m, 8H), 2
.78 (m, 2H), 3.2-3.4 (m, 5H), 3.54 (s, 3H), 6.41 (s, 1H), 7.10 (m, 1H), 7.2 (m
, 3H), 7.29 (d, 1H), 7.41 (dd, 1H), 7.71 (dd, 1H), 8.49 (d, 1H), 8.89 (dd, 1H
.), 9.91 (br S, 1H) FAB MS:. 558 (M + + H), 580 (M + + Na).

G. Synthesis of Compound 7 (+)-1,2,3,6-tetrahydro-1- {N- [4- (2-pyridyl) piperidin-1-yl] propyl} carboxamido-5-methoxy Carbonyl-4-hydroxymethyl-6- (3,4-
Difluorophenyl) -2-oxopyrimidine a) (+)-1,2,3,6-tetrahydro-1- {N- [4- (2-pyridyl) piperidin-1-yl]
Propyl} carboxamido-5-methoxycarbonyl-4-hydroxymethyl-6-
(+)-1,2,3,6-tetrahydro-1- {N- [4- (2-pyridyl) piperidine-1- (3,4-difluorophenyl) -2-oxopyrimidine in dichloromethane (20 mL) Yl] propyl} carboxamido-5-methoxycarbonyl
-4-methoxymethyl-6- (3,4-difluorophenyl) -2-oxopyrimidine (250 ml
, 0.448 mmol) (as described in ID (d)) was added dropwise at −78 ° C. to boron tribromide (1.0 M in dichloromethane, 1.20 mL, 1.20 mmol). The cooling bath was removed and the reaction mixture was gradually warmed to room temperature with stirring, and after 4 h MeOH (5 mL
) And concentrated to give a residue. Residue in chloroform / MeOH / 2N in MeOH
Purification by preparative thin-layer chromatography using ammonia = 100: 4: 1 as eluent gave the desired product (15 mg, 6%). MS: (M + H) 544. ¹ H-NMR (300 MHZ, CDCl ₃ )
δ1.79-1.98 (m, 6H), 2.05-2.12 (m, 2H), 2.45 (t, J = 7 Hz, 2H), 2.68-2.76 (m,
1H), 3.04-3.08 (m, 2H), 3.25-3.42 (m, 2H), 3.68 (s, 3H), 4.95 (dd, J = 17 Hz,
23 Hz, 2H), 6.65 (s, 1H), 6.97-7.23 (m, 5H), 7.62 (t, J 8 Hz, 1H), 8.48-8.
50 (d, J = 5 Hz, 1H), 8.98 (t, J = 7 Hz, 1H).

H. Synthesis of Compound 8 (+)-1-3-{[4- (3,4-difluorophenyl) -2,5-dioxo-1,2,5,7-tetrahydro-4H-furo [ 3,4-d] -pyrimidine-3-carbonyl] amino} -propyl-4- (2-pyridyl) -piperidine a) Methyl 2-{(3,4-difluorophenyl) methylene} -3-oxobutyrate 3,4-Difluorobenzaldehyde (14.2 g, 0.1 mol) in benzene (150 ml)
, Methyl acetoacetate (12.2 g, 0.105 mol), piperidine (0.430 g, 5 mmol), and acetic acid (0.30 g, 5 mmol) were stirred and refluxed in a Dean-Stark trap for 8 hours. . The benzene was evaporated, the residue dissolved in ethyl acetate (200 mL), brine (50 mL), saturated potassium bisulfate solution (50 mL).
) And saturated sodium bicarbonate solution. The ethyl acetate solution was dried (magnesium sulfate), the solvent was removed under reduced pressure and the residue was purified by column chromatography (SiO2, EtOAc / hexane, 10-15%). The product methyl 2-{(3,4-difluorophenyl) methylene} -3-oxobutyrate was obtained as a yellow oil (0.98 g, 98.3%) which was purified without further identification. Used for step.

B) 6- (3,4-Difluorophenyl) -1,6-dihydro-2-methoxy-5-methoxycarbonyl-4-methylpyrimidine Methyl 2-{(3,4-methylpyrimidine in DMF (30 mL) (Difluorophenyl) methylene} -3-oxobutyrate (8.8 g, 36.6 mmol), O-methylisourea hydrogen sulfate (9.4 g, 55
Mmol), and a mixture of NaHCO ₃ (12.3 g, 0.146 mol) was stirred at 70 ° C. for 1 hour.
Heated for 6 hours. The mixture was cooled, diluted with EtOAc (300 mL), and water (5 x 300 mL)
Washed with brine (300 mL) and dried (MgSO ₄ ). The solvent was evaporated and the crude product was purified by flash column chromatography on silica gel using 10% to 20% EtOAc in hexane as a gradient eluent to leave the product as an oil (3.82 g, 30.2%). ¹ H-NMR (CDCl ₃ ): δ2.32, 2.39 (2s, 3H), 3.58,
3.64 (2s, 3H), 3.72, 3.85 (2s, 3H), 5.55 (s, 1H), 6.13-7.8 (m, 4H).

C) 6- (3,4-Difluorophenyl) -1,6-dihydro-2-methoxy-5-methoxycarbonyl-4-methyl-1-[(4-nitrophenyloxy) -carbonyl] pyrimidine CH 6- (3,4-Difluorophenyl) 1,6-dihydro-2-methoxy in ₂ Cl ₂ (50 mL)
-5-Methoxycarbonyl-4-methylpyrimidine (2.82 g, 9.52 mmol) and
To a solution of 4-dimethylaminopyridine (1.16 g, 9.52 mmol) was added 4-nitrophenyl chloroformate (1.82 g, 9.04 mmol) at 0-5 ° C and the mixture was allowed to warm to room temperature. After 12 hours the solvent was evaporated and the residue was purified by flash column chromatography (SiO _2, EtOAc / hexane, 10% to 15%) to give the product as white crystals (3.72, 84.7); mp 172 ~ 174 ° C. ¹ H-NMR (CDCl ₃ ): δ2
.51 (s, 3H), 3.72 (s, 3H), 3.97 (s, 3H), 6.26 (s, 1H), 7.0-7.3 (m, 3H), 7.38 (
d, J = 9.3 Hz, 2H), 8.32 (d J = 9.3 Hz, 2H).

D) (+)-6- (3,4-difluorophenyl) -1,6-dihydro-2-oxo-5-methoxy
-Carbonyl-4-bromomethyl-1-[(4-nitrophenyl-oxy) -carbonyl]
Pyrimidine (+)-6- (3,4-difluorophenyl) -1,6-dihydro-2-in 5 mL of chloroform
Methoxy-5-methoxycarbonyl-4-methyl-1-[(4-nitrophenyloxy)-
Carbonyl] To a well stirred solution of pyrimidine (1.5 mmol, 0.66 g) was added a solution of bromine (1.5 mmol, 0.09 mL) in 3 mL of chloroform at 0 ° C.
Allowed to reach room temperature over time. The solvent was removed in vacuo and the residue re-CHCl ₃ (2
0 mL) and washed with brine. The organic layer was separated, dried over Na ₂ SO _4, filtered and the solvent removed in vacuo to a yellow foam (+) - 6- (3,4-difluorophenyl) -1,
0.81 g of 6-dihydro-2-oxo-5-methoxycarbonyl-4-bromomethyl-1-[(4-nitrophenyloxy) carbonyl] pyrimidine was obtained. This was used in the next step without further purification. ¹ H NMR δ 3.75 (s, 3H), 4.67 (ABq, d _A = 4.
56, d _B = 4.78, J = 10.8 Hz, 2H), 6.35 (s, 1H), 7.09-7.19 (m, 4H), 7.37 (d, J =
90Hz, 2H), 8.27 (d, J = 9.0 Hz, 2H).

E) (+)-4- (3,4-difluoro-phenyl) -2,5-dioxo-1,2,4,5,6,7-hexahydro-cyclopentapyrimidine-3-carboxylic acid- Nitrophenyl ester (+)-6- (3,4-difluorophenyl) -1,6-dihydro-2-oxo-5-methoxy-
Carbonyl-4-bromomethyl-1-[(4-nitrophenyloxy) carbonyl] pyrimidine (1.5 mmol, 0.81 g) was heated in an oil bath for 3 hours (oil bath temperature 130 ° C.).
The brown residue obtained here was washed with CHCl ₃ to give (+)-4- (3,4-
Difluoro-phenyl) -2,5-dioxo-1,2,4,5,6,7-hexahydro-cyclopentapyrimidine-3-carboxylic acid-nitrophenyl ester was obtained which was purified without further purification. (Crude substance weight 0.51 g). ¹ H NMR (DMSO)
δ4.94 (br, s, 2H), 6.08 (s, 1H), 7.20-7.43 (m, 4H), 8.35 (d, J = 10.2 Hz, 2H)
.

F) (+)-1-3-{[4- (3,4-difluorophenyl) -2,5-dioxo-1,2,5,7-tetrahydro-4H-furo [3,4- d] -Pyrimidine-3-carbonyl] amino} -propyl-
4- (2-pyridyl) -piperidine (+)-4- (3,4-difluoro-phenyl) -2,5-dioxo-1,2,4 in 10 mL of anhydrous THF
, 5,6,7-Hexahydro-cyclopentapyrimidine-3-carboxylic acid-4-nitrophenyl ester ((0.33 mmol, 0.14 g) and 3- [4- (2-pyridyl) -piperidine
A solution of -1-yl] propylamine (0.33 mmol, 0.07 g) was stirred at room temperature overnight. The solvent was removed in vacuo, the residue was purified by column chromatography (CH ₂ Cl _2, then CH ₂ C
l ₂ / MeOH 9: 1) to give the as an oil (+) - 1-3 - {[4- (3,4-difluorophenyl) -2,5-dioxo-1,2,5,7 Tetrahydro-4H-furo [3,4-d] -pyrimidine-3-carbonyl] amino} -propyl-4- (2-pyridyl) -piperidine was obtained (0.
16g, 98%). ¹ H NMR δ1.77-1.98 (m, 6H), 2.20 -2.25 (m, 2H), 2.54 (t, J = 7.2
Hz, 2H), 2.74-2.80 (m, 1H), 3.11-3.18 (m, 2H), 3.29-3.39 (m, 2H), 4.86 (s,
2H), 6.04 (br a, 1H), 6.46 (s, 1H), 7.07-7.26 (m, 5H), 7.63 (dt J = 7.8 Hz, J
= 1.8 Hz, 1H), 8.48 (d, J = 3.9 Hz), 9.22 (brt, J = 5.1 Hz, 1H). This was identified as dihydrochloride (hygroscopic). 90-95 ° C; [α] = +43.4 (c = 0.25, acetone);
_{Analysis: C 26 H 29 N 5 O} 4 F 2 C1 2 4.0H Calcd for _{2 O:. C, 47.57;} H, 5.68; N, 10.67 Found: C, 47.59; H, 5.32 ; N, 10.79.

I. Synthesis of Compound 9: 3- (3-carboxypropylcarbamoyl) -4 (S)-(3,4-difluorophenyl) -2-
Oxo-1,2,3,4-tetrahydro-4H-furo [3,4-d] pyrimidine a) (+)-5-methoxycarbonyl-4-methoxymethyl-1,2,3,6-tetrahydro-2
-Oxo-6 (S)-(3,4-difluorophenyl) -1-[(2-ethoxycarbonylethylamino) -carbonyl] pyrimidine β-alanine ethyl ester hydrochloride (87 mg, 0.57 mmol), (+)-
5-methoxycarbonyl-4-methoxymethyl-1,2,3,6-tetrahydro-2-oxo-
6 (S)-(3,4-difluorophenyl) -1-[(4-nitrophenyloxy) carbonyl]
Pyrimidine (as described in IA (c)) (227 mg, 0.48 mmol) and triethylamine (209 mL, 1.50 mmol) were combined in 5 mL of dichloromethane. The yellow mixture was stirred at ambient temperature for 1 hour and then evaporated to dryness. The resulting oil is
Purified by "flash" chromatography (5.02, dichloromethane: methanol: ammonium hydroxide 98: 2: 0.2). The product fractions were evaporated to dryness to give the title compound as a clear oil.

B) (+)-5-Methoxycarbonyl-4-methoxymethyl-1,2,3,6-tetrahydro-2
-Oxo-6 (S)-(3,4-difluorophenyl) -1-[(2-carboxyethylamino)-
Carbonyl] pyrimidine (+)-5-methoxycarbonyl-4-methoxymethyl-1,2,3,6-tetrahydro-2-oxo-6 (S)-(3,4-difluorophenyl) -1-[(2 -Ethoxycarbonylethylamino) carbonyl] pyrimidine (168 mg, 0.37 mmol) was dissolved in 5 mL of methanol. A 2.0 M aqueous solution of sodium hydroxide (400 mL, 0.80 mmol) was added dropwise. The mixture was stirred at ambient temperature for 4.5 hours, then concentrated in vacuo. The obtained residue was partitioned between ethyl acetate and 2.0 M aqueous hydrochloric acid. The phases were separated and the aqueous phase was extracted twice with ethyl acetate. The combined organic phases were then washed with brine. The organic phase was then dried over sodium sulfate, filtered and the filtrate was evaporated to dryness. The residue was re-concentrated from diethyl ether. The resulting white foamy solid was purified by "flash" chromatography (5.02
, Dichloromethane: methanol: glacial acetic acid 98: 2: 0.2). The product fractions were evaporated to dryness and the residue was re-concentrated from diethyl ether to give the title compound as a white foam solid. Mp: 52 ~55 ℃ NMR: Consistent with structure HPLC: purity 100% FAB MS: M + H @ m / e = 428.0 _{Analysis: C 18 H 19 F 2 N} 3 O 7 · 0.15H 2 O · 0.10Et 2 O Calculated for: C, 50.51; H, 4.6
8; N, 9.61. Found: C, 50.50; 15H, 4.46; N, 9.51.

C) 3- (3-carboxypropylcarbamoyl) -4 (S)-(3,4-difluorophenyl
) -2,5-Dioxo-1,2,3,4-tetrahydro-4H-furo [3,4-d] pyrimidine (+)-5-methoxycarbonyl-4-methoxymethyl-1,2,3,6 -Tetrahydro-2-
Dissolve oxo-6 (S)-(3,4-difluorophenyl) -1-[(2-carboxyethylamino) carbonyl] pyrimidine (308 mg, 0.72 mmol) in 5 mL of dichloromethane and dry under a nitrogen atmosphere while stirring. Cooled to -78 ° C in an ice / acetone bath. Then a 1.0 M solution of boron tribromide in dichloromethane (3.25 mL, 3.3 mmol) was added. The mixture was stirred at -78 ° C for 45 minutes, then gradually warmed to ambient temperature for 1 hour. 15 mL of saturated sodium bicarbonate was added until a neutral pH was reached.
The mixture was stirred at ambient temperature for 10 minutes. Volatile components were removed by evaporation.
The resulting aqueous solution was stirred at ambient temperature for 18 hours. The aqueous solution was then acidified with concentrated hydrochloric acid and the white solid that precipitated was filtered. The resulting white solid contains 0.1% TFA
H ₂ O: purified by preparative reverse-phase HPLC using CH ₃ CN gradient. The product fractions were evaporated to dryness to give the title compound as a white solid. NMR: consistent with structure HPLC: 96.2% pure FAB MS: M+H@m/e=382.1.

II. Synthesis of Other Potential Precursors Schemes 4A and 4B describe the synthesis of other potential precursors that can be used to make further compounds (Marquis et al., J. Med. Chem. . (1998) 41:
3563).

III. Oral Compositions As in certain embodiments relating to oral compositions of compounds of the present invention, 100 mg of one of the compounds described herein is formulated with sufficient fine lactose and
Or 590 mg total and filled into size 0 hard gel capsules.

IV. Pharmacological Properties of the Compounds at the Cloned Human Adrenergic Receptor The binding affinities of the selected compounds of the present invention were determined for the six cloned human α ₁ and α ₂ receptor subtypes, Measured on the channel. The protocols for these experiments are set forth below. 1. activity of the compound in a protocol various human receptor capacity measurement of alpha ₂ antagonist was measured in vitro using cultured cell lines that selectively express the receptor of interest. These cell lines were prepared by transfecting cloned cDNA or cloned genomic DNA or a construct containing both genomic DNA and cDNA, encoding the human α-adrenergic receptor as follows.

Α _1d human adrenergic receptor: 150 base pairs of 5 ′ untranslated sequence (5′UT)
And 300bp of 3 'untranslated sequence entire coding region of the alpha _1d containing (3'UT) (1719bp)
Is a eukaryotic expression vector pCEXV-3 modified with a polylinker called EXJ.HR
At the BamHI and ClaI sites. The construct required the joining of partially overlapping human lymphocyte genome and hippocampal cDNA clones. 5 'sequence is 1.2Kb
(The BamHI site from the vector was used for subcloning instead of the SmaI site from the internal insert), and the 3 'sequence was 1.3 Kb.
It was included in the XhoI-ClaI cDNA fragment (the ClaI site was derived from the vector polylinker). Using calcium phosphate technique, stabilized by co-transfection into LM (tk-) cells with plasmid α1A / EXJ (plasmid containing the aminoglycoside transferase gene) and the plasmid pGCcos3neo (α _1a receptor gene (expression vector containing the old name)) Cell line was obtained. Cells are controlled environment (37
C, 5% CO ₂ ) containing 25 mM glucose, 10% bovine serum, 100 units / ml
Were grown as monolayers in Dulbecco's modified Eagle's medium (GIBCO, Grand Island, NY) supplemented with Penicillin g and Streptomycin sulfate at 100 μg / ml.
Stable clones were then selected for resistance to the antibiotic G-418 (1 mg / ml) and membranes were harvested and assayed for their ability to bind [ ³ H] prazosin as described below (“Radioactive Ligand Binding Assay ").

As used herein, the cell line expressing the human _α1d receptor is designated L- _α1A (old name) and is the Budapest Treaty for the International Recognition of the Deposit of Microorganisms for the Purpose of Patent Procedure American Type Culture Collection, 12301 Park
deposited at lawn Drive, Rockville, Maryland 20852, USA. Cell line expressing the human α _1d receptor was deposited on September 25, 1992 as ATCC Accession No. CRL11138.

Α _1b human adrenergic receptor: 200p base pair 5 ′ untranslated sequence (5′UT
) And the entire coding region of the alpha _1b including 3 'untranslated sequence (3'UT) of 600bp (1563bp
) Was cloned into the EcoRI site of the pCESV-3 eukaryotic expression vector. The structure is
It was necessary to ligate the full length brain stem cDNA fragment derived from λZapII containing EcoRI to the expression vector. Stable cell lines were selected as described above. The cell line used herein is designated as L- _α1B and is subject to the provisions of the American Type Culture Collection, 1230 under the terms of the Budapest Treaty for the international recognition of the deposit of microorganisms for the purpose of patent procedures.
1 Deposited at Parklawn Drive, Rockville, Maryland 20852 USA. Cell line L
- α _1B it was observed on September 29, 1992 under ATCC Accession No. CR11139.

Α _1a human adrenergic receptor: 400 base pairs of 5 ′ untranslated sequence (5′UT)
And 200bp of 3 'untranslated sequence entire coding region of the alpha _1a containing (3'UT) (1401bp)
PCEXV-3 eukaryotic expression vector modified with polylinker, KpnI of EXJ.RH.
Was cloned into the site. The construct consists of three partially overlapping fragments: 5 '0.6kb
HincII genomic clone, middle 1.8 EcoRI hippocampus cDNA clone, and 3 '
Of the PstI genomic clone of 0.6 Kb. Since the hippocampal cDNA fragment overlaps with the 5 'and 3' genomic clones, HincII and PstI sites were used for ligation at the 5 'and 3' ends of the cDNA clone, respectively. The full-length clones were cloned into the vector (ie, pBluescript) and the 5 ′ and
It was cloned into the KpnI site of the expression vector using the 3'KpnI site. Stable cell lines were selected as described above. It represents a stable cell line expressing the human alpha _1a receptor used herein L-Arufa1c and (old name), the terms of the Budapest Treaty for the International Recognition of the Deposit of Microorganisms for the purpose of Patent Procedure American Typ
e Culture Collection, 12301 Parklawn Drive, Rockville, Maryland 20852 U.
Deposited with SA. Cell line expressing the human alpha _1a receptor with Accession No. CR11140 19
Approved on September 25, 1992.

Α₂ Receptor radioligand binding assay: Transfer from culture flask
Slice cells into 5 ml of 5 mM Tris-HCl, 5 mM EDTA, pH 7.5.
Dissolved by sonication. Centrifuge cell lysate at 1000 rpm for 5 minutes at 4 ° C
The supernatant was centrifuged at 30,000 xg for 20 minutes at 4 ° C. Pellet pH 7.5
50 mM Tris-HCl, 1 mM MgCl_Two And 0.1% ascorbic acid. α
₂ Antagonists [^ThreeH] Prazosin (0.5 nM, specific activity 76.2 Ci / mmol) LM (t
k-) Binding of the cells to the membrane preparation was performed in a final volume of 0.25 ml and incubation at 37 ° C for 20 minutes.
Incubated. Non-specific binding was measured in the presence of 10 μM phentolamine
. The reaction is performed by filtering through a GF / B filter using a cell harvester.
Stopped. Inhibition experiments routinely consist of 7 concentrations of test compound, which
Analysis was performed using a regression curve fitting computer program to obtain Ki values.

[0124] alpha ₂ Human Adrenergic Receptor: To measure the ability of in alpha ₂ receptor alpha ₁ antagonists, were stably transfected with the gene encoding alpha _2a, the alpha _2b and alpha _2c receptor The LM (tk-) cell line was used. The cell line expressing the α2a receptor is expressed as L-alpha _2A, it was deposited on November 6, 1992 under ATCC Accession No. CRL11180. cell line expressing the alpha _2b receptor represents the L-NGC- α _2B, was deposited on October 25, 1989 under ATCC Accession No. CRL10275. cell lines expressing the α _2c receptor represents a L- α _2c, was deposited on November 6, 1992 under ATCC Accession No. CRL-11181. All cell lines were obtained under the terms of the American Type Culture Collection, 1230, under the terms of the Budapest Treaty for the international recognition of the deposit of microorganisms for the purpose of patent procedures.
1 Deposited at Parklawn Drive, Rockville, Maryland 20852 USA. Cell lysates were prepared as described above (see radiobinding assay) and suspended in 25 mM glycylglycine buffer (pH 7.6 at room temperature). The equilibrium competitive binding assay was performed with [3H] rauwolscine (0.5 nM) and non-specific binding was determined by incubation with 10 μM phentolamine. Bound radioligand was separated by filtration through a GF / B filter using a cell harvester.

[0125] Measurement of the activity of alpha ₂ antagonist calcium channel: the ability of calcium channel alpha ₁ antagonists, essentially Glossma
n and Ferry (Methods in Enzymology 109: 513-550, 1985) as determined by a competitive binding assay of [3H] nitrendipine for membrane fragments of rat myocardium. For convenience, the tissue is minced and homogenized in 50 mM Tris-HCl (pH 7.4) containing 0.1 mM phenylmethylsulfonyl fluoride. Centrifuge the homogenate at 1000g for 15 minutes and centrifuge the resulting supernatant at 45,000g for 15 minutes. Resuspend the 45,000 g pellet in buffer and centrifuge a second time. An aliquot of the membrane protein was then removed at 37 ° C in the presence of [3H] nitrendipine (1 nM).
Incubate for 30 minutes at, and measure non-specific binding in the presence of 10 μM nifedipine. Bound radioligand is separated by filtration through a GF / B filter using a cell harvester.

The above compounds were assayed using the cloned human alpha adrenergic receptor. Preferred compounds that are selective α _1a antagonists have been found. The binding affinities of several compounds are illustrated in the table below.

Binding affinity of compounds of the invention selected at the cloned human α _1d , α _1b , α _1a , α _2a , α _2b and α _2c receptors (h = human).

[0128]

[Table 1]

[0129]

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[0130]

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[0131]

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[0132]

Embedded image

[0133]

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[0134]

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[0135]

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[0136]

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[0137]

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[0138]

[Brief description of the drawings]

FIG. 1A shows the structures of compounds 1 to 4 described herein in the Examples.

FIG. 1B shows the structures of compounds 5-8 described herein in the Examples.

FIG. 1C shows the structure of Compound 9 described herein in the Examples.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) A61K 31/56 A61K 31/56 45/00 45/00 A61P 13/08 A61P 13/08 C07D 401/12 C07D 401/12 401/14 401/14 491/048 491/048 (81) Designated country EP (AT, BE, CH, CY, DE, DK, ES, FI, FR, GB, GR, IE, IT, LU , MC, NL, PT, SE), AU, CA, JP (72) Inventor Kui, Dongguan, United States, 18940 Newtown, PA, Brownsberg Road West 280 (72) Inventor Davis, Margaret Earl 98053 Redmond, Washington, USA NE Seventy Six Place 20636 (72) Invention Kali, Hanumarpie, United States, Pennsylvania 19440 Hatfield, Heartford Drive 1064 (72) Inventor Biers, Kamreshpie, United States of America, Pennsylvania 19454 North Wales, Kent Drive 113 (72) Inventor Jean Evans, Benjamin United States of America, Pennsylvania 19446 Landsdale, Parkiomen Ave 501 (72) Inventor Dan, Michael United States of America, Mass. 02138 Cambridge, CA 92129 San Diego, Senda Panacea 12631 (72) Box 222, Oxford Street 12 (72) Inventor Naglas Nam, Danaparan 07446 Ramsey, Island Avenue 450, Apa, New Jersey, United States 07446 Arrangement 74 (72) Inventor Lagu, Barrat New Jersey, United States 07607 Maywood, East Pleasant Avenue 94 F-term (reference) 4C050 AA01 BB08 CC16 EE02 FF05 GG03 HH04 4C063 AA01 AA03 BB09 CC31 DD10 DD12 EE01 4C076 AA11A36 FF11 4C084 AA19 MA02 MA16 MA34 MA35 NA05 ZA811 ZC202 4C086 AA01 AA02 AA03 BC42 CB22 GA07 GA08 MA01 MA02 MA04 MA16 MA34 MA35 NA14 ZA81

Claims (33)

[Claims] 1. The following structure: Wherein R ₁ is —OCH ₃ or OH; R ₂ is —CH ₂ OH, —CH ₂ OCH ₃ or —COOH; R ₁ and R ₂ together form a 5-membered lactone ring; R ₃ is-(CH ₂ ) ₃ OH, Selected from the group consisting of: (where R ₁ is OH, R ₃ is Or a pharmaceutically acceptable salt thereof. 2. The (-) enantiomer of the compound according to claim 1. 3. The (+) enantiomer of the compound according to claim 1. 4. The following structure: The compound of claim 1 having the formula: 5. The following structure: 5. The compound according to claim 4, having the formula: 6. The following structure: 5. The compound according to claim 4, having the formula: 7. The following structure: 5. The compound according to claim 4, having the formula: 8. The following structure: embedded image 5. The compound according to claim 4, having the formula: (9) R ₃ is The compound of claim 4, wherein the compound is selected from the group consisting of: 10. The following structure: embedded image The compound of claim 1 having the formula: 11. The following structure: embedded image A compound according to claim 10 having the formula: 12. The following structure: A compound according to claim 10 having the formula: 13. The following structure: A compound according to claim 10 having the formula: 14. The following structure: A compound having the formula: 15. A pharmaceutical composition comprising a therapeutically effective amount of a compound according to claim 1, 6, 8, 9, 12, 13 or 14 and a pharmaceutically acceptable carrier. 16. The therapeutically effective amount is from about 0.01 mg to about 500 mg.
A pharmaceutical composition according to claim 15. 17. The pharmaceutical composition according to claim 16, wherein the therapeutically effective amount is from about 0.1 mg to about 60 mg. 18. The pharmaceutical composition according to claim 17, wherein the therapeutically effective amount is from about 1 mg to about 30 mg. 19. The method according to claim 15, wherein the carrier is a liquid and the composition is a solution.
A pharmaceutical composition according to claim 1. 20. The method of claim 15, wherein the carrier is a solid and the composition is a tablet.
A pharmaceutical composition according to claim 1. 21. The method according to claim 15, wherein the carrier is a gel and the composition is a suppository.
A pharmaceutical composition according to claim 1. 22. The method according to claim 1, which is effective for treating benign prostatic hyperplasia.
A method of treating a patient suffering from benign prostatic hyperplasia, comprising administering to a patient an amount of the compound of claim 8. 23. The method according to claim 1, which is effective for treating benign prostatic hyperplasia.
A method for treating a patient suffering from benign prostatic hyperplasia, comprising administering to a patient an amount of the compound of claim 5 in combination with a 5α-reductase inhibitor. 24. The 5-alpha reductase inhibitor is finasteride.
3. The method according to 3. 25. A method of relaxing sub-urinary tissue comprising administering to a patient an amount of a compound of claim 1 effective to relax sub-urinary tissue. 26. The method of claim 25, wherein the lower urinary tract tissue is urethral smooth muscle. 27. A method of inhibiting contraction of prostate tissue in a patient, comprising administering an amount of a compound according to claim 1 or 14 effective to inhibit contraction of prostate tissue. 28. A disease susceptible to treatment by antagonism of the _α1A receptor, comprising administering to a patient an amount of a compound according to claim 1 or 14 which is effective to treat the disease. How to treat. 29. A pharmaceutical composition comprising a therapeutically effective amount of a compound of claim 1 or 14 in combination with a therapeutically effective amount of finasteride and a pharmaceutically acceptable carrier. 30. The pharmaceutical composition according to claim 29, wherein the amount of the compound in a therapeutically effective amount is from about 0.01 mg to about 500 mg, and the amount of the therapeutically effective finasteride is about 5 mg. 31. The pharmaceutical composition according to claim 29, wherein the amount of the therapeutically effective compound is from about 0.1 mg to about 60 mg, and the amount of the therapeutically effective finasteride is about 5 mg. 32. The pharmaceutical composition according to claim 29, wherein the amount of the therapeutically effective compound is from about 1 mg to about 30 mg, and the amount of the therapeutically effective finasteride is about 5 mg. 33. A method for producing a pharmaceutical composition comprising combining a therapeutically effective amount of a compound of claim 1 or 14 with a pharmaceutically acceptable carrier.