JPWO2016152759A1 - 6-Aza-tetrahydro-γ-carboline derivative and pharmaceutical use thereof - Google Patents

Abstract

An object of the present invention is to provide a compound having a 5-HT2C receptor agonist activity and useful as a therapeutic or preventive agent for stress urinary incontinence. The present invention provides a 6-aza-tetrahydro-γ-carboline derivative represented by the following chemical formula or a pharmaceutically acceptable acid addition salt thereof.

Description

  The present invention relates to 6-aza-tetrahydro-γ-carboline derivatives and pharmaceutical uses thereof.

  Stress urinary incontinence means that urine leaks when transient abdominal pressure rises in daily life, such as when sneezing or coughing, laughing, running or lifting a heavy object It is a disease characterized by symptoms. The cause is thought to be due to trauma, ischemia or denervation of the pelvic floor muscle group or external urethral sphincter, which is a urethral closure tissue, due to childbirth, menopause or aging, etc. Many have been reported (Non-Patent Document 1).

  Treatment of stress urinary incontinence includes exercise therapy, surgery, or drug therapy. Pelvic floor muscle training, which is exercise therapy, can be effective after several weeks to several months by performing it continuously every day (Non-patent Document 2). No (non-patent document 3). Surgical surgery has been shown to have a certain effect, but since it depends on the severity and the will of the patient, there are currently few patients who actually undergo surgery (Non-patent Document 4).

  Regarding drug therapy, only duloxetine and clenbuterol are clinically used as therapeutic agents for the indication of stress urinary incontinence. Duloxetine contracts the urethra by suppressing the reuptake of serotonin and noradrenaline at the nerve endings, thereby providing a therapeutic effect. Clenbuterol stimulates the adrenergic β2 receptor of the external urethral sphincter and contracts the urethra to produce a therapeutic effect. However, the treatment rate and the treatment continuation rate with these drugs are low (Non-patent Document 5), and the development of a new therapeutic agent or preventive agent for stress urinary incontinence is eagerly desired.

  In recent years, it has been proposed that there is a urinary forbidden reflex mechanism that promotes urethral contraction when transient abdominal pressure rises (Non-Patent Document 6), and activates 5-HT2C receptor, which is one of serotonin receptors. It has been reported that a compound, that is, a 5-HT2C receptor agonist, can act as a therapeutic or preventive agent for stress urinary incontinence by promoting the urinary forbidden reflex mechanism (Patent Documents 1 to 3 and Non-Patent Documents). Reference 7).

  On the other hand, as the 6-aza-tetrahydro-γ-carboline derivative, for example, a compound described in Patent Document 4 has been reported.

JP 2012-107032 A International Publication No. 2011-111817 International Publication No. 2008/117169 International Publication No. 2010/080253

Irwin et al., British Journal of Urology Int. 2011, Vol. 108, No. 7, p. 1132-1139 Sampselle et al., Journal of Obstetric, Gynecologic, and Neonatal Nursing, 1997, Vol. 26, No. 4, p. 375-385 Bishop et al., Journal of Obstetric, Gynecologic, and Neonatal Nursing, 1992, Vol. 21, No. 5, p. 401-406 Waetjen et al., OBSTETRICS & GYNECOLOGY, 2003, 101, No. 4, p. 671-676 Vella et al., International Urology Journal and Pelic Floor Dysfunction, 2008, Vol. 19, No. 7, p. 961-964 Yoshimura et al., Japanese Pharmacology Journal, 2003, Vol. 121, No. 5, p. 290-298 Miyazaki et al., American Journal of Physiology. Renal physology, 2009, Vol. 297, No. 4, p. F1024-1031

  However, Patent Documents 1 to 4 and Non-Patent Document 7 do not describe that 6-aza-tetrahydro-γ-carboline derivatives have 5-HT2C receptor agonist activity, and even suggest the possibility thereof. Not.

  Therefore, an object of the present invention is to provide a compound having 5-HT2C receptor agonist activity and useful as a therapeutic or preventive agent for stress urinary incontinence.

  As a result of intensive studies to achieve the above object, the present inventors have found that a novel 6-aza-tetrahydro-γ-carboline derivative or a pharmaceutically acceptable acid addition salt thereof is a potent 5-HT2C. The present inventors have found that it has receptor agonist activity and have completed the present invention.

［式中、Ｒ^１は、炭素数１〜６のアルキル基、炭素数１若しくは２のアルキルチオ基、又は、ハロゲン原子、を表し、Ｒ^２は、水素原子、炭素数１〜３のアルキルオキシ基、又は、環構成原子数５若しくは６のヘテロアリール基、を表し、Ｒ^３は、水素原子又は炭素数１〜６のアルキル基を表し、Ｒ^１が炭素数１〜６のアルキル基又はハロゲン原子である場合、Ｒ^２は、炭素数１〜３のアルキルオキシ基又は環構成原子数５若しくは６のヘテロアリール基である。］That is, the present invention provides a 6-aza-tetrahydro-γ-carboline derivative represented by the following general formula (I) or a pharmaceutically acceptable acid addition salt thereof.

[Wherein, R ¹ represents an alkyl group having 1 to 6 carbon atoms, an alkylthio group having 1 or 2 carbon atoms, or a halogen atom, and R ² represents a hydrogen atom or an alkyloxy group having 1 to 3 carbon atoms. Or a heteroaryl group having 5 or 6 ring atoms, R ³ represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms, and R ¹ is an alkyl group or halogen atom having 1 to 6 carbon atoms. In this case, R ² is an alkyloxy group having 1 to 3 carbon atoms or a heteroaryl group having 5 or 6 ring atoms. ]

In the 6-aza-tetrahydro-γ-carboline derivative represented by the above general formula (I) or a pharmaceutically acceptable acid addition salt thereof, R ¹ is an alkyl group having 1 to 6 carbon atoms, 1 carbon atom or 2 is an alkylthio group or a chlorine atom, R ² is a hydrogen atom, an alkyloxy group having 1 to 3 carbon atoms, or a 3-thienyl group, and R ³ is preferably a hydrogen atom, R ¹ is an n-propyl group, R ² is a methoxy group or a 3-thienyl group, R ³ is more preferably a hydrogen atom, R ¹ is a methylthio group, and R ² is More preferably, a hydrogen atom or a methoxy group, and R ³ is a hydrogen atom.

  The present invention also provides a pharmaceutical and 5-HT2C receptor containing, as an active ingredient, a 6-aza-tetrahydro-γ-carboline derivative represented by the above general formula (I) or a pharmaceutically acceptable acid addition salt thereof. Provide body agonists.

  The medicament is preferably a therapeutic or prophylactic agent for stress urinary incontinence.

  The 6-aza-tetrahydro-γ-carboline derivative of the present invention and a pharmaceutically acceptable acid addition salt thereof have a potent 5-HT2C receptor agonistic activity, and have an excellent therapeutic effect on stress urinary incontinence Or exerts a preventive effect.

It is a figure which shows the effect | action of the compound of Example 1 with respect to the pressure at the time of rat leak under anesthesia.

［式中、Ｒ^１は、炭素数１〜６のアルキル基、炭素数１若しくは２のアルキルチオ基、又は、ハロゲン原子、を表し、Ｒ^２は、水素原子、炭素数１〜３のアルキルオキシ基、又は、環構成原子数５若しくは６のヘテロアリール基、を表し、Ｒ^３は、水素原子又は炭素数１〜６のアルキル基を表し、Ｒ^１が炭素数１〜６のアルキル基又はハロゲン原子である場合、Ｒ^２は、炭素数１〜３のアルキルオキシ基又は環構成原子数５若しくは６のヘテロアリール基である。］The 6-aza-tetrahydro-γ-carboline derivative of the present invention is characterized by being represented by the following general formula (I).

[Wherein, R ¹ represents an alkyl group having 1 to 6 carbon atoms, an alkylthio group having 1 or 2 carbon atoms, or a halogen atom, and R ² represents a hydrogen atom or an alkyloxy group having 1 to 3 carbon atoms. Or a heteroaryl group having 5 or 6 ring atoms, R ³ represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms, and R ¹ is an alkyl group or halogen atom having 1 to 6 carbon atoms. In this case, R ² is an alkyloxy group having 1 to 3 carbon atoms or a heteroaryl group having 5 or 6 ring atoms. ]

  “C1-C6 alkyl group” means a straight-chain saturated hydrocarbon group having 1 to 6 carbon atoms or a branched chain hydrocarbon group having 3 to 6 carbon atoms, such as a methyl group or an ethyl group. N-propyl group, isopropyl group, n-butyl group, isobutyl group, sec-butyl group, tert-butyl group, n-pentyl group or n-hexyl group.

  “Alkylthio group having 1 or 2 carbon atoms” means a methylthio group or an ethylthio group.

  “Halogen atom” means a fluorine atom, a chlorine atom, a bromine atom or an iodine atom.

  “Alkyloxy group having 1 to 3 carbon atoms” means a methoxy group, an ethoxy group, an n-propyloxy group, or an isopropyloxy group.

  The “heteroaryl group having 5 or 6 ring atoms” means the number of ring atoms containing 1 to 4 of the same or different atoms selected from the group consisting of a nitrogen atom, an oxygen atom and a sulfur atom. Means a heteroaromatic group of 5 or 6, for example, pyrrolyl group, furanyl group, thienyl group, pyrazolyl group, isoxazolyl group, imidazolyl group, oxazolyl group, thiazolyl group, pyridyl group, pyridyl group, pyrimidyl group or pyrazyl group Is mentioned.

In the 6-aza-tetrahydro-γ-carboline derivative represented by the above general formula (I), R ¹ is an alkyl group having 1 to 6 carbon atoms, an alkylthio group having 1 or 2 carbon atoms, or a chlorine atom. , R ² is a hydrogen atom, an alkyloxy group having 1 to 3 carbon atoms, or a 3-thienyl group, R ³ is preferably a hydrogen atom, and R ¹ is an n-propyl group, More preferably, R ² is a methoxy group or a 3-thienyl group, R ³ is a hydrogen atom, R ¹ is a methylthio group, R ² is a hydrogen atom or a methoxy group, and R ³ Is more preferably a hydrogen atom.

  Examples of the “pharmaceutically acceptable acid addition salt” of the 6-aza-tetrahydro-γ-carboline derivative represented by the above general formula (I) include, for example, hydrochloride, sulfate, nitrate, hydrobromide , Inorganic acid salts such as hydroiodide or phosphate, acetate, trifluoroacetate, lactate, citrate, oxalate, glutarate, malate, tartrate, fumarate, Organic carboxylates such as mandelate, maleate, benzoate or phthalate, or methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, camphorsulfonate, etc. Organic sulfonates are mentioned, but hydrochloride, hydrobromide, trifluoroacetate, tartrate or methanesulfonate is preferred, and hydrochloride or trifluoroacetate is more preferred. There.

  Among the 6-aza-tetrahydro-γ-carboline derivatives represented by the above general formula (I), preferred specific examples are shown in Table 1. However, these do not limit the present invention.

  The 6-aza-tetrahydro-γ-carboline derivative represented by the above general formula (I) can be produced by an appropriate method based on characteristics derived from the basic skeleton and the type of substituent. In addition, the compound which does not show the manufacturing method among the starting materials, a reagent, and a manufacturing intermediate used for this manufacture can generally be purchased.

［式中、Ｒ^１、Ｒ^２及びＲ^３は、上記定義に同じであり、Ｂｏｃは、ｔｅｒｔ−ブトキシカルボニル基を表す。］(Production method 1)
The 6-aza-tetrahydro-γ-carboline derivative represented by the above general formula (I) is, for example, 6-aza-tetrahydro represented by the general formula (II) in the presence of an acid as shown in the following scheme 1. -It can manufacture by the deprotection reaction of (gamma) -carboline derivative.

[Wherein R ¹ , R ² and R ³ are the same as defined above, and Boc represents a tert-butoxycarbonyl group. ]

  Examples of the acid used for the deprotection reaction include hydrogen chloride, sulfuric acid, p-toluenesulfonic acid, and trifluoroacetic acid. Hydrogen chloride or trifluoroacetic acid is preferable, and hydrogen chloride is more preferable.

  The equivalent amount of the acid used for the deprotection reaction is preferably 5 to 1000 equivalents, more preferably 10 to 400 equivalents, relative to the 6-aza-tetrahydro-γ-carboline derivative represented by the general formula (II).

  Examples of the solvent used for the deprotection reaction include tetrahydrofuran (hereinafter referred to as THF), 1,4-dioxane, acetonitrile, dichloromethane, methanol, or ethanol, and 1,4-dioxane, dichloromethane, or methanol is preferable. 4-dioxane is more preferred.

  The concentration at the start of the reaction of the 6-aza-tetrahydro-γ-carboline derivative represented by the above general formula (II) in the deprotection reaction is preferably 0.1 mmol / L to 5 mol / L, and 1 mmol / L to 1 mol. / L is more preferable.

  0-60 degreeC is preferable and, as for the reaction temperature of deprotection reaction, 20-40 degreeC is more preferable.

  The reaction time for the deprotection reaction is appropriately selected depending on the reaction temperature and other conditions, but is preferably 10 minutes to 72 hours, more preferably 30 minutes to 24 hours.

［式中、Ｂｏｃは、上記定義に同じであり、Ｘは、臭素原子又は塩素原子を表し、ＰＧは、保護基を表し、Ｒ^４は、メチル基又はエチル基を表す。］(Production method 2)
Among the 6-aza-tetrahydro-γ-carboline derivatives represented by the above general formula (II), the compound (II-a) can be produced in three steps as shown in the following scheme 2, for example. .

[Wherein Boc is the same as defined above, X represents a bromine atom or a chlorine atom, PG represents a protecting group, and R ⁴ represents a methyl group or an ethyl group. ]

(Process 1)
The 6-aza-tetrahydro-γ-carboline derivative (IV) can be produced, for example, by introducing a protecting group into the 6-aza-tetrahydro-γ-carboline derivative (V).
The introduction of the protecting group varies depending on the kind of the protecting group, and is performed, for example, according to the method described in Protective Groups in Organic Synthesis 3rd edition (Green et al., 1999, John Wiley & Sons, Inc.) or a method analogous thereto. be able to.

(Process 2)
The 6-aza-tetrahydro-γ-carboline derivative (III) is produced, for example, by thioetherification of the above 6-aza-tetrahydro-γ-carboline derivative (IV) with sodium thioalkoxide (LI). be able to.

  The equivalent of the sodium thioalkoxide (LI) used in the thioetherification reaction is preferably 1 to 10 equivalents, and 1.1 to 5 equivalents relative to the 6-aza-tetrahydro-γ-carboline derivative (IV). Is more preferable.

  Examples of the solvent used in the thioetherification reaction include THF, 1,4-dioxane, N, N-dimethylformamide (hereinafter referred to as DMF) or N, N-dimethylacetamide (hereinafter referred to as DMA), and DMF or DMA. Is preferred, and DMA is more preferred.

  The concentration of the 6-aza-tetrahydro-γ-carboline derivative (IV) at the start of the reaction in the thioetherification reaction is preferably 0.1 mmol / L to 10 mol / L, more preferably 0.01 to 1 mol / L. .

  0-110 degreeC is preferable and, as for the reaction temperature of thioetherification reaction, 60-100 degreeC is more preferable.

  The reaction time of the thioetherification reaction is appropriately selected according to conditions such as reaction temperature, but is preferably 10 minutes to 72 hours, more preferably 30 minutes to 48 hours.

(Process 3)
The 6-aza-tetrahydro-γ-carboline derivative (II-a) can be produced, for example, by deprotecting the 6-aza-tetrahydro-γ-carboline derivative (III).

  The deprotection of the protecting group varies depending on the type of the protecting group. For example, the protecting group is protected according to the method described in Protective Groups in Organic Synthesis 3rd edition (Green et al., 1999, John Wiley & Sons, Inc.) or a method similar thereto. It can be carried out.

［式中、Ｘ及びＢｏｃは、上記定義に同じである。］(Production method 3)
The 6-aza-tetrahydro-γ-carboline derivative (V) can be produced in two steps as shown in Scheme 3 below, for example.

[Wherein, X and Boc are the same as defined above. ]

(Process 1)
The hydrazonopiperidine derivative (VI) can be produced, for example, by a hydrazone formation reaction between a hydrazinylpyridine derivative (VII) and tert-butyl 4-oxopiperidine-1-carboxylate (VIII).

  The equivalent of the tert-butyl 4-oxopiperidine-1-carboxylate (VIII) used in the hydrazone formation reaction is preferably 1 to 3 equivalents relative to the hydrazinylpyridine derivative (VII). 5 equivalents are more preferred.

  Examples of the solvent used for the hydrazone formation reaction include THF, 1,4-dioxane, toluene, methanol, and ethanol, and methanol or ethanol is preferable.

  The concentration of the hydrazinylpyridine derivative (VII) at the start of the reaction in the hydrazone formation reaction is preferably 0.1 mmol / L to 10 mol / L, more preferably 0.1 to 1 mol / L.

  0-60 degreeC is preferable and, as for the reaction temperature of hydrazone formation reaction, 20-40 degreeC is more preferable.

  The reaction time of the hydrazone formation reaction is appropriately selected according to the reaction temperature and other conditions, but is preferably 10 minutes to 72 hours, more preferably 30 minutes to 48 hours.

(Process 2)
The 6-aza-tetrahydro-γ-carboline derivative (V) can be produced, for example, by Boc protection reaction in the presence of a base after the indole formation reaction of the hydrazonopiperidine derivative (VI) in the presence of an acid. it can.

  Examples of the acid used for the indole formation reaction include hydrogen chloride, sulfuric acid, p-toluenesulfonic acid, phosphoric acid, polyphosphoric acid, and acetic acid. Hydrogen chloride or polyphosphoric acid is preferable, and polyphosphoric acid is more preferable.

  As for the equivalent of the acid used for indole formation reaction, 2-1000 equivalent is preferable with respect to said hydrazono piperidine derivative (VI), and 5-400 equivalent is more preferable.

  Examples of the solvent used for the indole formation reaction include THF, 1,4-dioxane, acetonitrile, and toluene, and toluene is preferable.

  In the indole formation reaction, the concentration of the hydrazonopiperidine derivative (VI) at the start of the reaction is preferably 0.1 mmol / L to 5 mol / L, and more preferably 0.01 to 1 mol / L.

  20-200 degreeC is preferable and, as for the reaction temperature of indole formation reaction, 80-180 degreeC is more preferable.

  The reaction time of the indole formation reaction is appropriately selected according to conditions such as the reaction temperature, but is preferably 1 hour to 72 hours, and more preferably 3 hours to 48 hours.

  Examples of the base used for the Boc protection reaction include sodium hydride, tert-butoxy sodium, tert-butoxy potassium, sodium hydroxide, triethylamine, sodium carbonate, or potassium carbonate, and sodium carbonate or potassium carbonate is preferable.

  1-10 equivalent is preferable with respect to said hydrazono piperidine derivative (VI), and, as for the equivalent of the base used for Boc protection reaction, 1.5-5 equivalent is more preferable.

The equivalent of di-tert-butyl dicarbonate (hereinafter referred to as (Boc) ₂ O) used in the Boc protection reaction is preferably 1 to 5 equivalents relative to the hydrazonopiperidine derivative (VI), and 1.1 to 3 equivalents. Is more preferable.

  Examples of the solvent used for the Boc protection reaction include THF, 1,4-dioxane, dichloromethane, DMF, methanol, ethanol, isopropanol, and water. Isopropanol or water is preferable, and these may be used in combination. .

  In the Boc protection reaction, the concentration of the hydrazonopiperidine derivative (VI) at the start of the reaction is preferably 0.1 mmol / L to 5 mol / L, and more preferably 0.01 to 1 mol / L.

  0-60 degreeC is preferable and the reaction temperature of Boc protection reaction has more preferable 20-40 degreeC.

  The reaction time of the Boc protection reaction is appropriately selected according to the reaction temperature and other conditions, but is preferably 10 minutes to 72 hours, more preferably 30 minutes to 24 hours.

  The pharmaceutically acceptable acid addition salt of the 6-aza-tetrahydro-γ-carboline derivative represented by the above general formula (I) is, for example, 6-aza-tetrahydro- represented by the above general formula (I). It can be produced by a chlorination reaction by mixing a γ-carboline derivative and an acid.

  Examples of the acid used for the chlorination reaction include inorganic acids such as hydrogen chloride, sulfuric acid, nitric acid, hydrobromic acid, hydroiodic acid, and phosphoric acid, acetic acid, trifluoroacetic acid, lactic acid, citric acid, oxalic acid, and glutaric acid. , Organic carboxylic acids such as malic acid, tartaric acid, fumaric acid, mandelic acid, maleic acid, benzoic acid or phthalic acid, or organic such as methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid or camphorsulfonic acid A sulfonic acid is mentioned.

  The chlorination reaction is generally performed in a solvent, and a solvent that does not inhibit the reaction is appropriately selected. Examples of such solvents include aliphatic alcohols such as methanol, ethanol or isopropanol, ethers such as diethyl ether, THF, 1,4-dioxane or ethylene glycol dimethyl ether, and amides such as DMF or N-methylpyrrolidone. , Sulfoxides such as dimethyl sulfoxide, aliphatic nitriles such as acetonitrile or propionitrile, ketones such as acetone or 2-butanone, esters such as ethyl acetate, methyl acetate or n-butyl acetate, or water. 1,4-dioxane is preferred.

  The medicament of the present invention and the 5-HT2C receptor agonist contain the 6-aza-tetrahydro-γ-carboline derivative represented by the above general formula (I) or a pharmaceutically acceptable acid addition salt thereof as an active ingredient. It is characterized by doing. The medicament is preferably a therapeutic or prophylactic agent for stress urinary incontinence.

  The 6-aza-tetrahydro-γ-carboline derivative represented by the above general formula (I) or a pharmaceutically acceptable acid addition salt thereof has 5-HT2C receptor agonist activity, for example, as described in Jerman et al. It can be confirmed in an in vitro experimental system using cells expressing human 5-HT2C receptor by a method according to the literature (European Journal of Pharmacology, 2001, Vol. 414, p. 23-30). Specifically, for example, human 5-HT2C receptor can be forcibly expressed in HEK-293 cells derived from human fetal kidney, and the increase in intracellular calcium concentration due to the agonistic activity of the compound can be measured.

  In addition, the 6-aza-tetrahydro-γ-carboline derivative represented by the above general formula (I) or a pharmaceutically acceptable acid addition salt thereof is effective in treating or preventing stress urinary incontinence, This can be confirmed by measuring a leak point pressure indicating urethral resistance in the urine accumulation period.

  Leak Point Pressure is the intravesical pressure when urine leakage occurs in the absence of detrusor contraction, and indicates the maximum urethral resistance that can resist the increase in intravesical pressure. In an animal model, it has been shown that a drug having an effect of increasing the leakage pressure is effective for treating or preventing stress urinary incontinence.

  As a method of measuring the leakage time pressure, for example, Lee et al. (International Urology Journal, 2003, Vol. 14, No. 1, p. 31-37), Miyazaki et al. (American Journal of Physiology. physiology, 2008, Vol. 295, No. 1, p.F264-271), and it is reported that duloxetine, a therapeutic agent for stress urinary incontinence, increases the pressure at the time of leakage. (Miyazato et al., American Journal of Physiology. 2008, 295, No. 1, p. F264-271).

  The 6-aza-tetrahydro-γ-carboline derivative represented by the above general formula (I) or a pharmaceutically acceptable acid addition salt thereof is a mammal (eg, mouse, rat, hamster, rabbit, dog, monkey, It can be used as a useful medicament (especially a therapeutic or preventive agent for stress urinary incontinence) when administered to cows, sheep or humans), particularly humans.

  When the 6-aza-tetrahydro-γ-carboline derivative represented by the above general formula (I) or a pharmaceutically acceptable acid addition salt thereof is used clinically as a pharmaceutical, the above general formula (I) The indicated 6-aza-tetrahydro-γ-carboline derivative or a pharmaceutically acceptable acid addition salt thereof may be used as it is, or an excipient, stabilizer, preservative, buffer, solubilizer, emulsifier. Additives such as diluents or tonicity agents may be mixed as appropriate. Moreover, said pharmaceutical can be manufactured by a normal method using these pharmaceutical carriers as appropriate. Examples of the above-mentioned pharmaceutical administration forms include oral preparations such as tablets, capsules, granules, powders or syrups, parenteral preparations such as inhalants, injections, suppositories or liquids, or topical administration. Examples include ointments, creams, patches, and the like. Further, it may be a known continuous preparation.

  The medicament preferably contains 0.00001 to 90% by weight of the 6-aza-tetrahydro-γ-carboline derivative represented by the above general formula (I) or a pharmaceutically acceptable acid addition salt thereof. More preferably, the content is 0.01 to 70% by weight. The dose is appropriately selected according to the patient's symptom, age and weight, and administration method. As an amount of the active ingredient for an adult, 0.1 μg to 1 g per day for an injection, 1 μg to 10 g for an oral formulation, In the case of a patch, 1 μg to 10 g is preferable and can be administered once or several times.

  Examples of the pharmaceutically acceptable carrier or diluent of the above-mentioned pharmaceutical include, for example, binders (syrup, gelatin, gum arabic, sorbitol, polyvinyl chloride, tragacanth, etc.), excipients (sugar, lactose, corn starch, calcium phosphate, Sorbitol or glycine) or a lubricant (magnesium stearate, polyethylene glycol, talc, silica, etc.).

  The above medicines may be used in combination with or in combination with other drugs in order to supplement or enhance the therapeutic or preventive effect or reduce the dose.

  EXAMPLES Hereinafter, although an Example is shown and this invention is explained in full detail, this invention is not limited to these.

  In the following description, the solvent name shown in the NMR data indicates the solvent used for the measurement. The 400 MHz NMR spectrum was measured using a Bruker AVANCE 400 type nuclear magnetic resonance apparatus (Bruker). The chemical shift is represented by δ (unit: ppm) based on tetramethylsilane, and the signals are s (single line), d (double line), t (triple line), m (multiple line), and brs (respectively). Wide). The ESI-MS spectrum was measured using LCMS-2010EV (Shimadzu Corporation). As the silica gel, TLC Silica gel 60 F254 (Merck KGaA) was used.

２−ブロモ−６−ヒドラジニルピリジン（１０ｇ、５３ｍｍｏｌ）のメタノール（１００ｍＬ）溶液に、ｔｅｒｔ−ブチル ４−オキソピペリジン−１−カルボキシレート（１１ｇ、５３ｍｍｏｌ）を室温で加え、室温で１６時間撹拌した。反応混合物を濃縮し、得られた濃縮物をシリカゲルカラムクロマトグラフィー（溶出液：酢酸エチル／ｎ−ヘキサン）にて精製することで、ｔｅｒｔ−ブチル ４−（２−（６−ブロモピリジン−２−イル）ヒドラゾノ）ピペリジン−１−カルボキシレートを灰色がかった白色固体として得た（１２ｇ、収率６２％）。
ESI-MS m/z 369 [M+H]⁺.Example 1 Synthesis of 2- (methylthio) -6,7,8,9-tetrahydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine hydrochloride:
[Step 1] Synthesis of tert-butyl 4- (2- (6-bromopyridin-2-yl) hydrazono) piperidine-1-carboxylate:

To a solution of 2-bromo-6-hydrazinylpyridine (10 g, 53 mmol) in methanol (100 mL), tert-butyl 4-oxopiperidine-1-carboxylate (11 g, 53 mmol) was added at room temperature and stirred at room temperature for 16 hours. did. The reaction mixture was concentrated, and the resulting concentrate was purified by silica gel column chromatography (eluent: ethyl acetate / n-hexane) to give tert-butyl 4- (2- (6-bromopyridin-2- Yl) hydrazono) piperidine-1-carboxylate was obtained as an off-white solid (12 g, 62% yield).
ESI-MS m / z 369 [M + H] ⁺ .

ｔｅｒｔ−ブチル ４−（２−（６−ブロモピリジン−２−イル）ヒドラゾノ）ピペリジン−１−カルボキシレート（５．０ｇ、１４ｍｍｏｌ）のトルエン（５０ｍＬ）溶液に、ポリリン酸（２５ｇ）を室温で加え、１６０℃で２４時間撹拌した。反応混合物を室温に冷却し、トルエンを除去した後（デカンテーション）、残渣にｐＨが９．０以上になるまで、炭酸ナトリウム水溶液を加えた。析出した固体をろ過し、室温で減圧乾燥することで中間体を得た。得られた中間体のイソプロパノール（１０ｍＬ）、水（１０ｍＬ）溶液に炭酸カリウム（５．５ｇ、４０ｍｍｏｌ）、（Ｂｏｃ）_２Ｏ（５．２ｇ、２４ｍｍｏｌ）を室温で加え、室温で３時間撹拌した。反応混合物を濃縮後、酢酸エチルで抽出し、硫酸ナトリウムで乾燥後、濃縮した。得られた濃縮物をシリカゲルカラムクロマトグラフィー（溶出液：酢酸エチル／ｎ−ヘキサン）にて精製することで、ｔｅｒｔ−ブチル ２−ブロモ−７，８−ジヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン−６（９Ｈ）−カルボキシレートを灰色がかった白色固体として得た（１．４ｇ、収率２９％）。
¹H NMR (400 MHz, DMSO-d₆) δ: 11.71 (1H, brs), 7.83 (1H, d, J = 8.0 Hz), 7.20 (1H, d, J = 8.0 Hz), 4.84-4.53 (2H, m), 3.73-3.68 (2H, m), 2.79-2.74 (2H, m), 1.43 (9H, s).
ESI-MS m/z 352 [M+H]⁺.[Step 2] Synthesis of tert-butyl 2-bromo-7,8-dihydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine-6 (9H) -carboxylate:

To a solution of tert-butyl 4- (2- (6-bromopyridin-2-yl) hydrazono) piperidine-1-carboxylate (5.0 g, 14 mmol) in toluene (50 mL) was added polyphosphoric acid (25 g) at room temperature. , And stirred at 160 ° C. for 24 hours. The reaction mixture was cooled to room temperature, toluene was removed (decantation), and an aqueous sodium carbonate solution was added to the residue until the pH reached 9.0 or higher. The precipitated solid was filtered and dried under reduced pressure at room temperature to obtain an intermediate. To a solution of the obtained intermediate in isopropanol (10 mL) and water (10 mL), potassium carbonate (5.5 g, 40 mmol) and (Boc) ₂ O (5.2 g, 24 mmol) were added at room temperature, and the mixture was stirred at room temperature for 3 hours. . The reaction mixture was concentrated, extracted with ethyl acetate, dried over sodium sulfate, and concentrated. The resulting concentrate was purified by silica gel column chromatography (eluent: ethyl acetate / n-hexane) to give tert-butyl 2-bromo-7,8-dihydro-5H-pyrrolo [2,3-b. : 4,5-c ′] dipyridine-6 (9H) -carboxylate was obtained as an off-white solid (1.4 g, 29% yield).
¹ H NMR (400 MHz, DMSO-d ₆ ) δ: 11.71 (1H, brs), 7.83 (1H, d, J = 8.0 Hz), 7.20 (1H, d, J = 8.0 Hz), 4.84-4.53 (2H , m), 3.73-3.68 (2H, m), 2.79-2.74 (2H, m), 1.43 (9H, s).
ESI-MS m / z 352 [M + H] ⁺ .

ｔｅｒｔ−ブチル ２−ブロモ−７，８−ジヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン−６（９Ｈ）−カルボキシレート（５００ｍｇ、１．４ｍｍｏｌ）のＤＭＦ（３．０ｍＬ）溶液に、水素化ナトリウム（１１０ｍｇ、２．８ｍｍｏｌ）を０℃で加え、０℃で１０分間撹拌した後、２−（トリメチルシリル）エトキシメチルクロリド（０．３６ｍＬ、２．１ｍｍｏｌ）を０℃で加え、室温で１時間撹拌した。反応混合物に水（１０ｍＬ）を加え、酢酸エチル（２５ｍＬ×２）で抽出した。有機層をあわせ、硫酸ナトリウムで乾燥後、濃縮した。得られた濃縮物をシリカゲルカラムクロマトグラフィー（溶出液：メタノール／ジクロロメタン）にて精製することで、ｔｅｒｔ−ブチル ２−ブロモ−９−（（２−（トリメチルシリル）エトキシ）メチル）−７，８−ジヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン−６（９Ｈ）−カルボキシレートを白色固体として得た（５４０ｍｇ、収率７９％）。
ESI-MS m/z 482 [M+H]⁺.[Step 3] tert-Butyl 2-bromo-9-((2- (trimethylsilyl) ethoxy) methyl) -7,8-dihydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine- Synthesis of 6 (9H) -carboxylate:

tert-Butyl 2-bromo-7,8-dihydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine-6 (9H) -carboxylate (500 mg, 1.4 mmol) in DMF (3 0.0 mL) solution, sodium hydride (110 mg, 2.8 mmol) was added at 0 ° C. and stirred at 0 ° C. for 10 minutes, and then 2- (trimethylsilyl) ethoxymethyl chloride (0.36 mL, 2.1 mmol) was added to 0 The mixture was added at ° C and stirred at room temperature for 1 hour. Water (10 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (25 mL × 2). The organic layers were combined, dried over sodium sulfate, and concentrated. The resulting concentrate was purified by silica gel column chromatography (eluent: methanol / dichloromethane), whereby tert-butyl 2-bromo-9-((2- (trimethylsilyl) ethoxy) methyl) -7,8- Dihydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine-6 (9H) -carboxylate was obtained as a white solid (540 mg, 79% yield).
ESI-MS m / z 482 [M + H] ⁺ .

ｔｅｒｔ−ブチル ２−ブロモ−９−（（２−（トリメチルシリル）エトキシ）メチル）−７，８−ジヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン−６（９Ｈ）−カルボキシレート（３００ｍｇ、０．６２ｍｍｏｌ）のＤＭＡ（１．０ｍＬ）溶液に、ナトリウムチオメトキシド（１３０ｍｇ、１．９ｍｍｏｌ）を室温で加え、封管し、８０℃で３０分間撹拌した。反応混合物にジクロロメタン（３０ｍＬ）を室温で加え、水（１０ｍＬ×２）、飽和食塩水（１０ｍＬ）で順次洗浄した。有機層を硫酸ナトリウムで乾燥後、濃縮した。得られた濃縮物をシリカゲルカラムクロマトグラフィー（溶出液：１５％酢酸エチル／ｎ−ヘキサン）にて精製することで、ｔｅｒｔ−ブチル ２−（メチルチオ）−９−（（２−（トリメチルシリル）エトキシ）メチル）−７，８−ジヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン−６（９Ｈ）−カルボキシレートをガム状固体として得た（３００ｍｇ、収率９０％）。
¹H NMR (400 MHz, CDCl₃) δ: 7.55 (1H, d, J = 8.2 Hz), 6.96 (1H, d, J = 8.2 Hz), 5.59 (2H, s), 4.57 (2H, brs), 3.84 (2H, brs), 3.52 (2H, t, J= 8.2 Hz), 2.95 (2H, brs), 2.62 (3H, s), 1.50 (9H, s), 0.90 (2H, t, J = 8.2 Hz), -0.07 (9H, s).
ESI-MS m/z 450 [M+H]⁺.[Step 4] tert-Butyl 2- (methylthio) -9-((2- (trimethylsilyl) ethoxy) methyl) -7,8-dihydro-5H-pyrrolo [2,3-b: 4,5-c ′] Synthesis of dipyridine-6 (9H) -carboxylate:

tert-Butyl 2-bromo-9-((2- (trimethylsilyl) ethoxy) methyl) -7,8-dihydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine-6 (9H) -Sodium thiomethoxide (130 mg, 1.9 mmol) was added to a solution of carboxylate (300 mg, 0.62 mmol) in DMA (1.0 mL) at room temperature, sealed, and stirred at 80 ° C for 30 minutes. Dichloromethane (30 mL) was added to the reaction mixture at room temperature, and the mixture was washed successively with water (10 mL × 2) and saturated brine (10 mL). The organic layer was dried over sodium sulfate and concentrated. The resulting concentrate was purified by silica gel column chromatography (eluent: 15% ethyl acetate / n-hexane) to give tert-butyl 2- (methylthio) -9-((2- (trimethylsilyl) ethoxy). Methyl) -7,8-dihydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine-6 (9H) -carboxylate was obtained as a gummy solid (300 mg, 90% yield). .
¹ H NMR (400 MHz, CDCl ₃ ) δ: 7.55 (1H, d, J = 8.2 Hz), 6.96 (1H, d, J = 8.2 Hz), 5.59 (2H, s), 4.57 (2H, brs), 3.84 (2H, brs), 3.52 (2H, t, J = 8.2 Hz), 2.95 (2H, brs), 2.62 (3H, s), 1.50 (9H, s), 0.90 (2H, t, J = 8.2 Hz ), -0.07 (9H, s).
ESI-MS m / z 450 [M + H] ⁺ .

ｔｅｒｔ−ブチル ２−（メチルチオ）−９−（（２−（トリメチルシリル）エトキシ）メチル）−７，８−ジヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン−６（９Ｈ）−カルボキシレート（３００ｍｇ、０．６６ｍｍｏｌ）のＴＨＦ（８．０ｍＬ）溶液に、テトラブチルアンモニウムフルオリド（以下、ＴＢＡＦ）（２ｍｏｌ／Ｌ ＴＨＦ溶液、４．０ｍＬ）を室温で加え、６０℃で５時間撹拌した。反応混合物に水を加え、酢酸エチル（２０ｍＬ）で抽出した。有機層を水（１０ｍＬ×２）で洗浄し、硫酸ナトリウムで乾燥後、濃縮した。得られた濃縮物をシリカゲルカラムクロマトグラフィー（溶出液：２５％酢酸エチル／ｎ−ヘキサン）にて精製することで、ｔｅｒｔ−ブチル ２−（メチルチオ）−７，８−ジヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン−６（９Ｈ）−カルボキシレートを灰色がかった白色固体として得た（１７０ｍｇ、収率８１％）。
¹H NMR (400 MHz, CDCl₃) δ: 8.37 (1H, brs), 7.58 (1H, d, J = 8.2 Hz), 6.96 (1H, d, J = 8.1 Hz), 4.57 (2H, brs), 3.80 (2H, brs), 2.81 (2H, brs), 2.59 (3H, s), 1.50 (9H, s).
ESI-MS m/z 320 [M+H]⁺.[Step 5] Synthesis of tert-butyl 2- (methylthio) -7,8-dihydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine-6 (9H) -carboxylate:

tert-Butyl 2- (methylthio) -9-((2- (trimethylsilyl) ethoxy) methyl) -7,8-dihydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine-6 ( 9H) -carboxylate (300 mg, 0.66 mmol) in THF (8.0 mL) was added tetrabutylammonium fluoride (hereinafter referred to as TBAF) (2 mol / L THF solution, 4.0 mL) at room temperature. For 5 hours. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate (20 mL). The organic layer was washed with water (10 mL × 2), dried over sodium sulfate, and concentrated. The obtained concentrate was purified by silica gel column chromatography (eluent: 25% ethyl acetate / n-hexane), whereby tert-butyl 2- (methylthio) -7,8-dihydro-5H-pyrrolo [2 , 3-b: 4,5-c ′] dipyridine-6 (9H) -carboxylate was obtained as an off-white solid (170 mg, 81% yield).
¹ H NMR (400 MHz, CDCl ₃ ) δ: 8.37 (1H, brs), 7.58 (1H, d, J = 8.2 Hz), 6.96 (1H, d, J = 8.1 Hz), 4.57 (2H, brs), 3.80 (2H, brs), 2.81 (2H, brs), 2.59 (3H, s), 1.50 (9H, s).
ESI-MS m / z 320 [M + H] ⁺ .

ｔｅｒｔ−ブチル ２−（メチルチオ）−７，８−ジヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン−６（９Ｈ）−カルボキシレート（１７０ｍｇ、０．５４ｍｍｏｌ）に、４規定塩化水素１，４−ジオキサン溶液（３．６ｍＬ）を室温で加え、室温で２時間撹拌した。反応混合物を濃縮し、濃縮残渣にアセトニトリルを室温で加えた。析出した固体をろ取し、室温で減圧乾燥することで、２−（メチルチオ）−６，７，８，９−テトラヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン塩酸塩（以下、実施例１の化合物）を灰色がかった白色固体として得た（１２５ｍｇ、収率９１％）。
¹H NMR (400 MHz, DMSO-d₆) δ: 11.66 (1H, brs), 9.47 (2H, brs), 7.78 (1H, d, J = 8.2 Hz), 6.96 (1H, d, J = 8.2 Hz), 4.25 (2H, brs), 3.49-3.40 (2H, m), 2.99 (2H, t, J = 6.0 Hz), 2.53 (3H, s).
ESI-MS m/z 220 [M+H]⁺.[Step 6] Synthesis of 2- (methylthio) -6,7,8,9-tetrahydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine hydrochloride:

To tert-butyl 2- (methylthio) -7,8-dihydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine-6 (9H) -carboxylate (170 mg, 0.54 mmol), A 4N hydrogen chloride 1,4-dioxane solution (3.6 mL) was added at room temperature, and the mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated, and acetonitrile was added to the concentrated residue at room temperature. The precipitated solid was collected by filtration and dried under reduced pressure at room temperature to give 2- (methylthio) -6,7,8,9-tetrahydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine. The hydrochloride salt (hereinafter, the compound of Example 1) was obtained as an off-white solid (125 mg, 91% yield).
¹ H NMR (400 MHz, DMSO-d ₆ ) δ: 11.66 (1H, brs), 9.47 (2H, brs), 7.78 (1H, d, J = 8.2 Hz), 6.96 (1H, d, J = 8.2 Hz ), 4.25 (2H, brs), 3.49-3.40 (2H, m), 2.99 (2H, t, J = 6.0 Hz), 2.53 (3H, s).
ESI-MS m / z 220 [M + H] ⁺ .

ｔｅｒｔ−ブチル ２−（メチルチオ）−７，８−ジヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン−６（９Ｈ）−カルボキシレート（５０ｍｇ、０．１５ｍｍｏｌ）のＤＭＦ（２．５ｍＬ）溶液に、水素化ナトリウム（１２ｍｇ、０．２９ｍｍｏｌ）を０℃で加え、０℃で２０分間撹拌した。続いて、ヨウ化メチル（０．０２５ｍＬ、０．３９ｍｍｏｌ）を０℃で加え、室温で１時間撹拌した。反応混合物に氷水を加え、１０分間撹拌した後、酢酸エチル（１０ｍＬ×２）で抽出した。有機層をあわせ、飽和食塩水（５ｍＬ×２）で洗浄し、硫酸ナトリウムで乾燥後、濃縮した。得られた濃縮物をシリカゲルカラムクロマトグラフィー（溶出液：１０％酢酸エチル／ｎ−ヘキサン）にて精製することで、ｔｅｒｔ−ブチル ９−メチル−２−（メチルチオ）−７，８−ジヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン−６（９Ｈ）−カルボキシレートを灰色がかった白色固体として得た（２５ｍｇ、収率８９％）。
ESI-MS m/z 334 [M+H]⁺.Example 2 Synthesis of 9-methyl-2- (methylthio) -6,7,8,9-tetrahydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine hydrochloride:
[Step 1] tert-butyl 9-methyl-2- (methylthio) -7,8-dihydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine-6 (9H) -carboxylate Synthesis:

DMF of tert-butyl 2- (methylthio) -7,8-dihydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine-6 (9H) -carboxylate (50 mg, 0.15 mmol) To the (2.5 mL) solution, sodium hydride (12 mg, 0.29 mmol) was added at 0 ° C. and stirred at 0 ° C. for 20 minutes. Subsequently, methyl iodide (0.025 mL, 0.39 mmol) was added at 0 ° C., and the mixture was stirred at room temperature for 1 hour. Ice water was added to the reaction mixture, and the mixture was stirred for 10 minutes, and then extracted with ethyl acetate (10 mL × 2). The organic layers were combined, washed with saturated brine (5 mL × 2), dried over sodium sulfate, and concentrated. The obtained concentrate was purified by silica gel column chromatography (eluent: 10% ethyl acetate / n-hexane) to give tert-butyl 9-methyl-2- (methylthio) -7,8-dihydro-5H. -Pyrrolo [2,3-b: 4,5-c '] dipyridine-6 (9H) -carboxylate was obtained as an off-white solid (25 mg, 89% yield).
ESI-MS m / z 334 [M + H] ⁺ .

ｔｅｒｔ−ブチル ９−メチル−２−（メチルチオ）−７，８−ジヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン−６（９Ｈ）−カルボキシレート（２０ｍｇ、０．０６０ｍｍｏｌ）に、４規定塩化水素１，４−ジオキサン溶液（０．４０ｍＬ）を室温で加え、室温で２時間撹拌した。反応混合物を濃縮し、濃縮残渣にアセトニトリルを室温で加えた。析出した固体をろ取し、室温で減圧乾燥することで、９−メチル−２−（メチルチオ）−６，７，８，９−テトラヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン塩酸塩（以下、実施例２の化合物）を灰色固体として得た（１１ｍｇ、収率６８％）。
¹H NMR (400 MHz, DMSO-d₆) δ: 9.45 (2H, brs), 7.81 (1H, d, J = 8.2 Hz), 6.91 (1H, d, J = 8.2 Hz), 4.28 (2H, brs), 3.69 (3H, s), 3.54-3.46 (2H, m), 3.07 (2H, t, J= 5.9 Hz), 2.58 (3H, s).
ESI-MS m/z 234 [M+H]⁺.[Step 2] Synthesis of 9-methyl-2- (methylthio) -6,7,8,9-tetrahydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine hydrochloride:

tert-Butyl 9-methyl-2- (methylthio) -7,8-dihydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine-6 (9H) -carboxylate (20 mg, 0. 060 mmol), 4N hydrogen chloride 1,4-dioxane solution (0.40 mL) was added at room temperature, and the mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated, and acetonitrile was added to the concentrated residue at room temperature. The precipitated solid was collected by filtration and dried under reduced pressure at room temperature, whereby 9-methyl-2- (methylthio) -6,7,8,9-tetrahydro-5H-pyrrolo [2,3-b: 4,5- c ′] dipyridine hydrochloride (compound of Example 2 hereinafter) was obtained as a gray solid (11 mg, yield 68%).
¹ H NMR (400 MHz, DMSO-d ₆ ) δ: 9.45 (2H, brs), 7.81 (1H, d, J = 8.2 Hz), 6.91 (1H, d, J = 8.2 Hz), 4.28 (2H, brs ), 3.69 (3H, s), 3.54-3.46 (2H, m), 3.07 (2H, t, J = 5.9 Hz), 2.58 (3H, s).
ESI-MS m / z 234 [M + H] ⁺ .

水素化ナトリウム（２１０ｍｇ、５．３ｍｍｏｌ）のＴＨＦ（２．０ｍＬ）溶液に、無水メタノール（０．２０ｍＬ）を０℃で加え、０℃で２時間撹拌した。この反応混合物に、２，６−ジブロモ−４−ニトロピリジン（１．０ｇ、３．５ｍｍｏｌ）のＴＨＦ（３．０ｍＬ）溶液を０℃で加え、室温で３時間撹拌した。反応混合物に飽和塩化アンモニウム水溶液（１０ｍＬ）を加え、酢酸エチル（１００ｍＬ×２）で抽出した。有機層をあわせ、硫酸ナトリウムで乾燥後、濃縮した。得られた濃縮物を再結晶（酢酸エチル／ｎ−ヘキサン）にて精製することで、２，６−ジブロモ−４−メトキシピリジンを白色固体として得た（６３０ｍｇ、収率６６％）。
¹H NMR (400 MHz, CDCl₃) δ: 6.98 (2H, s), 3.85 (3H, s).
ESI-MS m/z 265 [M+H]⁺.Example 3 Synthesis of 4-methoxy-2-propyl-6,7,8,9-tetrahydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine hydrochloride:
[Step 1] Synthesis of 2,6-dibromo-4-methoxypyridine:

To a solution of sodium hydride (210 mg, 5.3 mmol) in THF (2.0 mL) was added anhydrous methanol (0.20 mL) at 0 ° C., and the mixture was stirred at 0 ° C. for 2 hours. To this reaction mixture, a solution of 2,6-dibromo-4-nitropyridine (1.0 g, 3.5 mmol) in THF (3.0 mL) was added at 0 ° C., and the mixture was stirred at room temperature for 3 hours. A saturated aqueous ammonium chloride solution (10 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (100 mL × 2). The organic layers were combined, dried over sodium sulfate, and concentrated. The obtained concentrate was purified by recrystallization (ethyl acetate / n-hexane) to obtain 2,6-dibromo-4-methoxypyridine as a white solid (630 mg, yield 66%).
¹ H NMR (400 MHz, CDCl ₃ ) δ: 6.98 (2H, s), 3.85 (3H, s).
ESI-MS m / z 265 [M + H] ⁺ .

２，６−ジブロモ−４−メトキシピリジン（５３０ｍｇ、２．０ｍｍｏｌ）のイソプロパノール（１０ｍＬ）溶液に、ヒドラジン一水和物（０．５０ｍＬ、１６ｍｍｏｌ）を室温で加え、１１０℃で２４時間撹拌した。反応混合物を室温に冷却し、濃縮後、濃縮残渣をクロロホルムに溶解した。この反応混合物にｎ−ヘキサンを加え、析出した固体をろ過し、室温で減圧乾燥することで、２−ブロモ−６−ヒドラジニル−４−メトキシピリジンを灰色がかった白色固体として得た（３７０ｍｇ、収率８４％）。
¹H NMR (400 MHz, CDCl₃) δ: 6.44 (1H, d, J = 2.0 Hz), 6.21 (1H, d, J = 2.0 Hz), 5.93 (1H, brs), 3.79 (3H, s), 3.75 (2H, brs).
ESI-MS m/z 217 [M+H]⁺.[Step 2] Synthesis of 2-bromo-6-hydrazinyl-4-methoxypyridine:

To a solution of 2,6-dibromo-4-methoxypyridine (530 mg, 2.0 mmol) in isopropanol (10 mL) was added hydrazine monohydrate (0.50 mL, 16 mmol) at room temperature, and the mixture was stirred at 110 ° C. for 24 hours. The reaction mixture was cooled to room temperature and concentrated, and then the concentrated residue was dissolved in chloroform. N-Hexane was added to the reaction mixture, and the precipitated solid was filtered and dried under reduced pressure at room temperature to obtain 2-bromo-6-hydrazinyl-4-methoxypyridine as an off-white solid (370 mg, yield). 84%).
¹ H NMR (400 MHz, CDCl ₃ ) δ: 6.44 (1H, d, J = 2.0 Hz), 6.21 (1H, d, J = 2.0 Hz), 5.93 (1H, brs), 3.79 (3H, s), 3.75 (2H, brs).
ESI-MS m / z 217 [M + H] ⁺ .

２−ブロモ−６−ヒドラジニル−４−メトキシピリジン（３５０ｍｇ、１．６ｍｍｏｌ）のエタノール（８．０ｍＬ）溶液に、ｔｅｒｔ−ブチル ４−オキソピペリジン−１−カルボキシレート（３５０ｍｇ、１．７ｍｍｏｌ）を室温で加え、室温で１時間撹拌した。反応混合物を濃縮し、得られた濃縮物をシリカゲルカラムクロマトグラフィー（溶出液：酢酸エチル／ｎ−ヘキサン）にて精製することで、ｔｅｒｔ−ブチル ４−（２−（６−ブロモ−４−メトキシピリジン−２−イル）ヒドラゾノ）ピペリジン−１−カルボキシレートを黄色オイルとして得た（６２０ｍｇ、収率９８％）。
¹H NMR (400 MHz, CDCl₃) δ: 7.76 (1H, s), 6.63 (1H, d, J = 2.0 Hz), 3.84 (3H, s), 3.56 (4H, t, J = 6.2 Hz), 2.50 (2H, t, J = 6.2 Hz), 2.37 (2H, t, J = 6.0 Hz), 1.48 (9H, s).
ESI-MS m/z 399 [M+H]⁺.[Step 3] Synthesis of tert-butyl 4- (2- (6-bromo-4-methoxypyridin-2-yl) hydrazono) piperidine-1-carboxylate:

To a solution of 2-bromo-6-hydrazinyl-4-methoxypyridine (350 mg, 1.6 mmol) in ethanol (8.0 mL) was added tert-butyl 4-oxopiperidine-1-carboxylate (350 mg, 1.7 mmol) at room temperature. And stirred at room temperature for 1 hour. The reaction mixture was concentrated, and the resulting concentrate was purified by silica gel column chromatography (eluent: ethyl acetate / n-hexane) to give tert-butyl 4- (2- (6-bromo-4-methoxy). Pyridin-2-yl) hydrazono) piperidine-1-carboxylate was obtained as a yellow oil (620 mg, 98% yield).
¹ H NMR (400 MHz, CDCl ₃ ) δ: 7.76 (1H, s), 6.63 (1H, d, J = 2.0 Hz), 3.84 (3H, s), 3.56 (4H, t, J = 6.2 Hz), 2.50 (2H, t, J = 6.2 Hz), 2.37 (2H, t, J = 6.0 Hz), 1.48 (9H, s).
ESI-MS m / z 399 [M + H] ⁺ .

ｔｅｒｔ−ブチル ４−（２−（６−ブロモ−４−メトキシピリジン−２−イル）ヒドラゾノ）ピペリジン−１−カルボキシレート（２２０ｍｇ、０．５４ｍｍｏｌ）のトルエン（６．０ｍＬ）溶液に、ポリリン酸（３．０ｇ）を室温で加え、１６０℃で３６時間撹拌した。反応混合物を室温に冷却し、トルエンを除去した後（デカンテーション）、残渣にｐＨが９．０以上になるまで、炭酸ナトリウム水溶液を加えた。この反応混合物にイソプロパノール（１０ｍＬ）、（Ｂｏｃ）_２Ｏ（１８０ｍｇ、０．８１ｍｍｏｌ）を室温で加え、室温で１時間撹拌した。反応混合物を濃縮後、クロロホルム（５０ｍＬ）で抽出した。有機層を水で洗浄し、硫酸ナトリウムで乾燥後、濃縮した。得られた濃縮物をシリカゲルカラムクロマトグラフィー（溶出液：酢酸エチル／ｎ−ヘキサン）にて精製することで、ｔｅｒｔ−ブチル ２−ブロモ−４−メトキシ−７，８−ジヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン−６（９Ｈ）−カルボキシレートを白色固体として得た（６０ｍｇ、収率２９％）。
¹H NMR (400 MHz, CDCl₃) δ: 9.00 (1H, brs), 6.64 (1H, s), 4.69 (2H, brs), 3.92 (3H, s), 3.76 (2H, t, J = 6.4 Hz), 2.81 (2H, t, J= 6.4 Hz), 1.50 (9H, s).
ESI-MS m/z 382 [M+H]⁺. [Step 4] Synthesis of tert-butyl 2-bromo-4-methoxy-7,8-dihydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine-6 (9H) -carboxylate:

To a solution of tert-butyl 4- (2- (6-bromo-4-methoxypyridin-2-yl) hydrazono) piperidine-1-carboxylate (220 mg, 0.54 mmol) in toluene (6.0 mL) was added polyphosphoric acid ( 3.0 g) at room temperature and stirred at 160 ° C. for 36 hours. The reaction mixture was cooled to room temperature, toluene was removed (decantation), and an aqueous sodium carbonate solution was added to the residue until the pH reached 9.0 or higher. To this reaction mixture, isopropanol (10 mL) and (Boc) ₂ O (180 mg, 0.81 mmol) were added at room temperature, and the mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated and extracted with chloroform (50 mL). The organic layer was washed with water, dried over sodium sulfate and concentrated. The resulting concentrate was purified by silica gel column chromatography (eluent: ethyl acetate / n-hexane), whereby tert-butyl 2-bromo-4-methoxy-7,8-dihydro-5H-pyrrolo [2 , 3-b: 4,5-c ′] dipyridine-6 (9H) -carboxylate was obtained as a white solid (60 mg, 29% yield).
¹ H NMR (400 MHz, CDCl ₃ ) δ: 9.00 (1H, brs), 6.64 (1H, s), 4.69 (2H, brs), 3.92 (3H, s), 3.76 (2H, t, J = 6.4 Hz ), 2.81 (2H, t, J = 6.4 Hz), 1.50 (9H, s).
ESI-MS m / z 382 [M + H] ⁺ .

ｔｅｒｔ−ブチル ２−ブロモ−４−メトキシ−７，８−ジヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン−６（９Ｈ）−カルボキシレート（１００ｍｇ、０．２６ｍｍｏｌ）のトルエン（８．０ｍＬ）溶液に、ｎ−プロピルボロン酸（３４ｍｇ、０．３９ｍｍｏｌ）、酢酸パラジウム（６．０ｍｇ、０．０２６ｍｍｏｌ）、２規定炭酸ナトリウム水溶液（０．３９ｍＬ、０．７８ｍｍｏｌ）、トリシクロヘキシルホスフィン（１１ｍｇ、０．０３９ｍｍｏｌ）を室温で加え、１２０℃で２時間撹拌した。反応混合物を室温に冷却し、水（４０ｍＬ）を加え、ジクロロメタン（５０ｍＬ×２）で抽出した。有機層をあわせ、硫酸ナトリウムで乾燥後、濃縮した。得られた濃縮物をシリカゲルカラムクロマトグラフィー（溶出液：酢酸エチル／ｎ−ヘキサン）にて精製することで、ｔｅｒｔ−ブチル ４−メトキシ−２−プロピル−７，８−ジヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン−６（９Ｈ）−カルボキシレートを淡褐色固体として得た（４６ｍｇ、収率５１％）。
ESI-MS m/z 346 [M+H]⁺. [Step 5] Synthesis of tert-butyl 4-methoxy-2-propyl-7,8-dihydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine-6 (9H) -carboxylate:

tert-Butyl 2-bromo-4-methoxy-7,8-dihydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine-6 (9H) -carboxylate (100 mg, 0.26 mmol) In toluene (8.0 mL), n-propylboronic acid (34 mg, 0.39 mmol), palladium acetate (6.0 mg, 0.026 mmol), 2N aqueous sodium carbonate (0.39 mL, 0.78 mmol), Tricyclohexylphosphine (11 mg, 0.039 mmol) was added at room temperature and stirred at 120 ° C. for 2 hours. The reaction mixture was cooled to room temperature, water (40 mL) was added, and the mixture was extracted with dichloromethane (50 mL × 2). The organic layers were combined, dried over sodium sulfate, and concentrated. The obtained concentrate was purified by silica gel column chromatography (eluent: ethyl acetate / n-hexane), whereby tert-butyl 4-methoxy-2-propyl-7,8-dihydro-5H-pyrrolo [2 , 3-b: 4,5-c ′] dipyridine-6 (9H) -carboxylate was obtained as a light brown solid (46 mg, 51% yield).
ESI-MS m / z 346 [M + H] ⁺ .

ｔｅｒｔ−ブチル ４−メトキシ−２−プロピル−７，８−ジヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン−６（９Ｈ）−カルボキシレート（９．０ｍｇ、０．０２０ｍｍｏｌ）のメタノール（４．０ｍＬ）溶液に、４規定塩化水素１，４−ジオキサン溶液（０．１３ｍＬ）を室温で加え、室温で２時間撹拌した。反応混合物を濃縮し、濃縮残渣を水、アセトニトリルに溶解した後、凍結乾燥することで、４−メトキシ−２−プロピル−６，７，８，９−テトラヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン塩酸塩（以下、実施例３の化合物）を淡褐色固体として得た（７．１ｍｇ、収率９７％）。
¹H NMR (400 MHz, DMSO-d₆) δ: 12.32-11.98 (1H, m), 9.52-9.41 (2H, m), 6.84 (1H, brs), 4.31-4.26 (2H, brs), 4.01 (3H, s), 3.17-3.10 (2H, brs), 3.05-3.00 (2H, m), 2.89-2.77 (2H, m), 1.81-1.68 (2H, m), 0.92 (3H, t, J = 6.7 Hz).
ESI-MS m/z 246 [M+H]⁺.[Step 6] Synthesis of 4-methoxy-2-propyl-6,7,8,9-tetrahydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine hydrochloride:

tert-Butyl 4-methoxy-2-propyl-7,8-dihydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine-6 (9H) -carboxylate (9.0 mg, 0.0. 020 mmol) in methanol (4.0 mL) was added 4N hydrogen chloride 1,4-dioxane solution (0.13 mL) at room temperature, and the mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated, the concentrated residue was dissolved in water and acetonitrile, and then lyophilized to give 4-methoxy-2-propyl-6,7,8,9-tetrahydro-5H-pyrrolo [2,3-b. : 4,5-c ′] dipyridine hydrochloride (hereinafter, the compound of Example 3) was obtained as a light brown solid (7.1 mg, yield 97%).
¹ H NMR (400 MHz, DMSO-d ₆ ) δ: 12.32-11.98 (1H, m), 9.52-9.41 (2H, m), 6.84 (1H, brs), 4.31-4.26 (2H, brs), 4.01 ( 3H, s), 3.17-3.10 (2H, brs), 3.05-3.00 (2H, m), 2.89-2.77 (2H, m), 1.81-1.68 (2H, m), 0.92 (3H, t, J = 6.7 Hz).
ESI-MS m / z 246 [M + H] ⁺ .

ｔｅｒｔ−ブチル ２−ブロモ−４−メトキシ−７，８−ジヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン−６（９Ｈ）−カルボキシレート（９９ｍｇ、０．２６ｍｍｏｌ）のＤＭＡ（０．４２ｍＬ）溶液に、ナトリウムチオメトキシド（５５ｍｇ、０．８０ｍｍｏｌ）を室温で加え、封管し、８０℃で３０分間撹拌した。反応混合物にジクロロメタン（１５ｍＬ）を室温で加え、水（５ｍＬ×２）、飽和食塩水（５ｍＬ）で順次洗浄した。有機層を硫酸ナトリウムで乾燥後、濃縮した。得られた濃縮物をシリカゲルカラムクロマトグラフィー（溶出液：酢酸エチル／ｎ−ヘキサン）にて精製することで、ｔｅｒｔ−ブチル ４−メトキシ−２−（メチルチオ）−７，８−ジヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン−６（９Ｈ）−カルボキシレートを茶色固体として得た（１０ｍｇ、収率１１％）。
ESI-MS m/z 350 [M+H]⁺.Example 4 Synthesis of 4-methoxy-2- (methylthio) -6,7,8,9-tetrahydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine hydrochloride:
[Step 1] of tert-butyl 4-methoxy-2- (methylthio) -7,8-dihydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine-6 (9H) -carboxylate Synthesis:

tert-Butyl 2-bromo-4-methoxy-7,8-dihydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine-6 (9H) -carboxylate (99 mg, 0.26 mmol) Sodium methoxide (55 mg, 0.80 mmol) was added at room temperature to a solution of the solution in DMA (0.42 mL), sealed and stirred at 80 ° C. for 30 minutes. Dichloromethane (15 mL) was added to the reaction mixture at room temperature, and the mixture was washed successively with water (5 mL × 2) and saturated brine (5 mL). The organic layer was dried over sodium sulfate and concentrated. The obtained concentrate was purified by silica gel column chromatography (eluent: ethyl acetate / n-hexane) to give tert-butyl 4-methoxy-2- (methylthio) -7,8-dihydro-5H-pyrrolo. [2,3-b: 4,5-c ′] dipyridine-6 (9H) -carboxylate was obtained as a brown solid (10 mg, 11% yield).
ESI-MS m / z 350 [M + H] ⁺ .

ｔｅｒｔ−ブチル ４−メトキシ−２−（メチルチオ）−７，８−ジヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン−６（９Ｈ）−カルボキシレート（１５ｍｇ、０．０４４ｍｍｏｌ）に、４規定塩化水素１，４−ジオキサン溶液（０．２８ｍＬ）を室温で加え、室温で２時間撹拌した。反応混合物を濃縮し、濃縮残渣にアセトニトリルを室温で加えた。析出した固体をろ取し、室温で減圧乾燥することで、４−メトキシ−２−（メチルチオ）−６，７，８，９−テトラヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン塩酸塩（以下、実施例４の化合物）を茶色固体として得た（１２ｍｇ、収率９７％）。
¹H NMR (400 MHz, DMSO-d₆) δ: 11.58 (1H, brs), 9.04 (2H, brs), 6.54 (1H, s), 4.29 (2H, brs), 3.89 (3H, s), 3.48 (2H, brs), 2.93 (2H, t, J = 5.9 Hz), 2.53 (3H, s).
ESI-MS m/z 250 [M+H]⁺.[Step 2] Synthesis of 4-methoxy-2- (methylthio) -6,7,8,9-tetrahydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine hydrochloride:

tert-Butyl 4-methoxy-2- (methylthio) -7,8-dihydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine-6 (9H) -carboxylate (15 mg, .0. 044 mmol), 4N hydrogen chloride 1,4-dioxane solution (0.28 mL) was added at room temperature, and the mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated, and acetonitrile was added to the concentrated residue at room temperature. The precipitated solid was collected by filtration and dried under reduced pressure at room temperature, whereby 4-methoxy-2- (methylthio) -6,7,8,9-tetrahydro-5H-pyrrolo [2,3-b: 4,5- c ′] dipyridine hydrochloride (hereinafter, the compound of Example 4) was obtained as a brown solid (12 mg, 97% yield).
¹ H NMR (400 MHz, DMSO-d ₆ ) δ: 11.58 (1H, brs), 9.04 (2H, brs), 6.54 (1H, s), 4.29 (2H, brs), 3.89 (3H, s), 3.48 (2H, brs), 2.93 (2H, t, J = 5.9 Hz), 2.53 (3H, s).
ESI-MS m / z 250 [M + H] ⁺ .

２，４，６−トリクロロピリジン（１．０ｇ、５．５ｍｍｏｌ）のＴＨＦ（１０ｍＬ）溶液に、ｎ−ブチルリチウム（２．５ｍｏｌ／Ｌ ｎ−ヘキサン溶液、２．２ｍＬ）を−７８℃で加え、−７８℃で１時間撹拌した。この反応混合物に、トリメチルシリルクロリド（０．７０ｍＬ、５．５ｍｍｏｌ）を加え、−７８℃で１時間撹拌した。反応混合物に飽和塩化アンモニウム水溶液（１０ｍＬ）を加え、酢酸エチル（３０ｍＬ×２）で抽出した。有機層をあわせ、硫酸ナトリウムで乾燥後、濃縮した。得られた濃縮物をシリカゲルカラムクロマトグラフィー（溶出液：２％酢酸エチル／ｎ−ヘキサン）にて精製することで、２，４，６−トリクロロ−３−（トリメチルシリル）ピリジンを無色オイルとして得た（１．２ｇ、収率８７％）。
¹H NMR (400 MHz, CDCl₃) δ: 7.27 (1H, s), 0.51 (9H, s).
ESI-MS m/z 255 [M+H]⁺.Example 5: 2-Chloro-4- (thiophen-3-yl) -6,7,8,9-tetrahydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine hydrochloride Synthesis:
[Step 1] Synthesis of 2,4,6-trichloro-3- (trimethylsilyl) pyridine:

N-Butyllithium (2.5 mol / L n-hexane solution, 2.2 mL) was added to a THF (10 mL) solution of 2,4,6-trichloropyridine (1.0 g, 5.5 mmol) at −78 ° C. , And stirred at −78 ° C. for 1 hour. Trimethylsilyl chloride (0.70 mL, 5.5 mmol) was added to the reaction mixture, and the mixture was stirred at −78 ° C. for 1 hour. A saturated aqueous ammonium chloride solution (10 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (30 mL × 2). The organic layers were combined, dried over sodium sulfate, and concentrated. The resulting concentrate was purified by silica gel column chromatography (eluent: 2% ethyl acetate / n-hexane) to give 2,4,6-trichloro-3- (trimethylsilyl) pyridine as a colorless oil. (1.2 g, 87% yield).
¹ H NMR (400 MHz, CDCl ₃ ) δ: 7.27 (1H, s), 0.51 (9H, s).
ESI-MS m / z 255 [M + H] ⁺ .

２，４，６−トリクロロ−３−（トリメチルシリル）ピリジン（５８０ｍｇ、２．３ｍｍｏｌ）のＴＨＦ（１０ｍＬ）溶液に、ヒドラジン一水和物（０．１５ｍＬ、４．６ｍｍｏｌ）を室温で加え、５０℃で１６時間撹拌した。反応混合物を室温に冷却し、ＴＢＡＦ（１ｍｏｌ／Ｌ ＴＨＦ溶液、２．３ｍＬ）を加え、室温で２時間撹拌した。反応混合物に水（１０ｍＬ）を加え、酢酸エチル（３０ｍＬ）で抽出した。有機層を水（１０ｍＬ）で洗浄し、硫酸ナトリウムで乾燥後、濃縮した。得られた濃縮物をシリカゲルカラムクロマトグラフィー（溶出液：２０−２５％酢酸エチル／ｎ−ヘキサン）にて精製することで、２，４−ジクロロ−６−ヒドラジニルピリジンを無色オイルとして得た（１９０ｍｇ、収率４６％）。
¹H NMR (400 MHz, CDCl₃) δ: 8.23 (1H, s), 6.71 (1H, d, J = 1.2 Hz), 6.65 (1H, d, J = 1.6 Hz), 4.32 (2H, s).
ESI-MS m/z 179 [M+H]⁺.[Step 2] Synthesis of 2,4-dichloro-6-hydrazinylpyridine:

To a solution of 2,4,6-trichloro-3- (trimethylsilyl) pyridine (580 mg, 2.3 mmol) in THF (10 mL) was added hydrazine monohydrate (0.15 mL, 4.6 mmol) at room temperature, and 50 ° C. For 16 hours. The reaction mixture was cooled to room temperature, TBAF (1 mol / L THF solution, 2.3 mL) was added, and the mixture was stirred at room temperature for 2 hours. Water (10 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (30 mL). The organic layer was washed with water (10 mL), dried over sodium sulfate, and concentrated. The resulting concentrate was purified by silica gel column chromatography (eluent: 20-25% ethyl acetate / n-hexane) to give 2,4-dichloro-6-hydrazinylpyridine as a colorless oil. (190 mg, 46% yield).
¹ H NMR (400 MHz, CDCl ₃ ) δ: 8.23 (1H, s), 6.71 (1H, d, J = 1.2 Hz), 6.65 (1H, d, J = 1.6 Hz), 4.32 (2H, s).
ESI-MS m / z 179 [M + H] ⁺ .

２，４−ジクロロ−６−ヒドラジニルピリジン（１８０ｍｇ、１．０ｍｍｏｌ）のエタノール（５．０ｍＬ）溶液に、ｔｅｒｔ−ブチル ４−オキソピペリジン−１−カルボキシレート（２２０ｍｇ、１．１ｍｍｏｌ）を室温で加え、室温で１時間撹拌した。反応混合物を濃縮し、得られた濃縮物をシリカゲルカラムクロマトグラフィー（溶出液：２０−３０％酢酸エチル／ｎ−ヘキサン）にて精製することで、ｔｅｒｔ−ブチル ４−（２−（４，６−ジクロロピリジン−２−イル）ヒドラゾノ）ピペリジン−１−カルボキシレートを白色固体として得た（３４０ｍｇ、収率９４％）。
¹H NMR (400 MHz, CDCl₃) δ: 7.85 (1H, s), 7.13 (1H, d, J = 1.4 Hz), 6.76 (1H, d, J = 1.2 Hz), 3.62-3.57 (4H, m), 2.50 (2H, t, J = 6.4 Hz), 2.38 (2H, t, J = 6.0 Hz), 1.48 (9H, s).
ESI-MS m/z 360 [M+H]⁺.[Step 3] Synthesis of tert-butyl 4- (2- (4,6-dichloropyridin-2-yl) hydrazono) piperidine-1-carboxylate:

To a solution of 2,4-dichloro-6-hydrazinylpyridine (180 mg, 1.0 mmol) in ethanol (5.0 mL) was added tert-butyl 4-oxopiperidine-1-carboxylate (220 mg, 1.1 mmol) at room temperature. And stirred at room temperature for 1 hour. The reaction mixture was concentrated, and the resulting concentrate was purified by silica gel column chromatography (eluent: 20-30% ethyl acetate / n-hexane) to give tert-butyl 4- (2- (4,6 -Dichloropyridin-2-yl) hydrazono) piperidine-1-carboxylate was obtained as a white solid (340 mg, 94% yield).
¹ H NMR (400 MHz, CDCl ₃ ) δ: 7.85 (1H, s), 7.13 (1H, d, J = 1.4 Hz), 6.76 (1H, d, J = 1.2 Hz), 3.62-3.57 (4H, m ), 2.50 (2H, t, J = 6.4 Hz), 2.38 (2H, t, J = 6.0 Hz), 1.48 (9H, s).
ESI-MS m / z 360 [M + H] ⁺ .

ｔｅｒｔ−ブチル ４−（２−（４，６−ジクロロピリジン−２−イル）ヒドラゾノ）ピペリジン−１−カルボキシレート（９５ｍｇ、０．２７ｍｍｏｌ）のトルエン（３．０ｍＬ）溶液に、ポリリン酸（１．５ｇ）を室温で加え、１６０℃で３６時間撹拌した。反応混合物を室温に冷却し、トルエンを除去した後（デカンテーション）、残渣にｐＨが９．０以上になるまで、炭酸ナトリウム水溶液を加えた。この反応混合物にイソプロパノール（５．０ｍＬ）、（Ｂｏｃ）_２Ｏ（９１ｍｇ、０．４１ｍｍｏｌ）を室温で加え、室温で１時間撹拌した。反応混合物を濃縮後、クロロホルム（２５ｍＬ）で抽出した。有機層を水で洗浄し、硫酸ナトリウムで乾燥後、濃縮した。得られた濃縮物をシリカゲルカラムクロマトグラフィー（溶出液：１０−２０％酢酸エチル／ｎ−ヘキサン）にて精製することで、ｔｅｒｔ−ブチル ２，４−ジクロロ−７，８−ジヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン−６（９Ｈ）−カルボキシレートを白色固体として得た（１０ｍｇ、収率１１％）。
¹H NMR (400 MHz, CDCl₃) δ: 10.64 (1H, brs), 7.06 (1H, s), 4.83 (2H, brs), 3.80 (2H, t, J= 5.4 Hz), 2.93 (2H, t, J = 5.6 Hz), 1.48 (9H, s).
ESI-MS m/z 343 [M+H]⁺.[Step 4] Synthesis of tert-butyl 2,4-dichloro-7,8-dihydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine-6 (9H) -carboxylate:

To a solution of tert-butyl 4- (2- (4,6-dichloropyridin-2-yl) hydrazono) piperidine-1-carboxylate (95 mg, 0.27 mmol) in toluene (3.0 mL) was added polyphosphoric acid (1. 5 g) was added at room temperature and stirred at 160 ° C. for 36 hours. The reaction mixture was cooled to room temperature, toluene was removed (decantation), and an aqueous sodium carbonate solution was added to the residue until the pH reached 9.0 or higher. To this reaction mixture, isopropanol (5.0 mL) and (Boc) ₂ O (91 mg, 0.41 mmol) were added at room temperature, and the mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated and extracted with chloroform (25 mL). The organic layer was washed with water, dried over sodium sulfate and concentrated. The resulting concentrate was purified by silica gel column chromatography (eluent: 10-20% ethyl acetate / n-hexane) to give tert-butyl 2,4-dichloro-7,8-dihydro-5H-pyrrolo. [2,3-b: 4,5-c ′] dipyridine-6 (9H) -carboxylate was obtained as a white solid (10 mg, 11% yield).
¹ H NMR (400 MHz, CDCl ₃ ) δ: 10.64 (1H, brs), 7.06 (1H, s), 4.83 (2H, brs), 3.80 (2H, t, J = 5.4 Hz), 2.93 (2H, t , J = 5.6 Hz), 1.48 (9H, s).
ESI-MS m / z 343 [M + H] ⁺ .

ｔｅｒｔ−ブチル ２，４−ジクロロ−７，８−ジヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン−６（９Ｈ）−カルボキシレート（１００ｍｇ、０．２９ｍｍｏｌ）のトルエン（１０ｍＬ）溶液に、チオフェン−３−イル−ボロン酸（５６ｍｇ、０．４３ｍｍｏｌ）、酢酸パラジウム（６．０ｍｇ、０．０２９ｍｍｏｌ）、２規定炭酸ナトリウム水溶液（０．１０ｍＬ）、トリシクロヘキシルホスフィン（１２ｍｇ、０．０４４ｍｍｏｌ）を室温で加え、１００℃で１２時間撹拌した。反応混合物を水に注ぎ、酢酸エチル（２５ｍＬ×３）で抽出した。有機層をあわせ、硫酸マグネシウムで乾燥後、濃縮した。得られた濃縮物をシリカゲルカラムクロマトグラフィー（溶出液：酢酸エチル／ｎ−ヘキサン）にて精製することで、ｔｅｒｔ−ブチル ２−クロロ−４−（チオフェン−３−イル）−７，８−ジヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン−６（９Ｈ）−カルボキシレートを灰色がかった白色固体として得た（６０ｍｇ、収率５４％）。
ESI-MS m/z 390 [M+H]⁺. [Step 5] tert-Butyl 2-chloro-4- (thiophen-3-yl) -7,8-dihydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine-6 (9H) -Synthesis of carboxylates:

tert-Butyl 2,4-dichloro-7,8-dihydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine-6 (9H) -carboxylate (100 mg, 0.29 mmol) in toluene (10 mL) in solution, thiophen-3-yl-boronic acid (56 mg, 0.43 mmol), palladium acetate (6.0 mg, 0.029 mmol), 2N aqueous sodium carbonate (0.10 mL), tricyclohexylphosphine (12 mg , 0.044 mmol) at room temperature and stirred at 100 ° C. for 12 hours. The reaction mixture was poured into water and extracted with ethyl acetate (25 mL × 3). The organic layers were combined, dried over magnesium sulfate and concentrated. The obtained concentrate was purified by silica gel column chromatography (eluent: ethyl acetate / n-hexane) to give tert-butyl 2-chloro-4- (thiophen-3-yl) -7,8-dihydro. -5H-pyrrolo [2,3-b: 4,5-c '] dipyridine-6 (9H) -carboxylate was obtained as an off-white solid (60 mg, 54% yield).
ESI-MS m / z 390 [M + H] ⁺ .

ｔｅｒｔ−ブチル ２−クロロ−４−（チオフェン−３−イル）−７，８−ジヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン−６（９Ｈ）−カルボキシレート（３０ｍｇ、０．０７６ｍｍｏｌ）のジクロロメタン（４．０ｍＬ）溶液に、４規定塩化水素１，４−ジオキサン溶液（２．０ｍＬ）を室温で加え、室温で２時間撹拌した。反応混合物を濃縮し、濃縮残渣に酢酸エチル／ｎ−ヘキサンを室温で加えた。析出した固体をろ取し、室温で減圧乾燥することで、２−クロロ−４−（チオフェン−３−イル）−６，７，８，９−テトラヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン塩酸塩（以下、実施例５の化合物）を黄茶色固体として得た（１５ｍｇ、収率６８％）。
¹H NMR (400 MHz, DMSO-d₆) δ: 12.1 (1H, s), 9.22 (2H, brs), 7.85-7.37 (1H, m), 7.79-7.77 (1H, m), 7.38-7.37 (1H, m), 7.10 (1H, s), 3.95 (2H, brs), 3.50-3.47 (2H, m), 3.06 (2H, t, J = 5.2 Hz).
ESI-MS m/z 290 [M+H]⁺.[Step 6] Synthesis of 2-chloro-4- (thiophen-3-yl) -6,7,8,9-tetrahydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine hydrochloride :

tert-butyl 2-chloro-4- (thiophen-3-yl) -7,8-dihydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine-6 (9H) -carboxylate ( To a solution of 30 mg, 0.076 mmol) in dichloromethane (4.0 mL) was added 4N hydrogen chloride 1,4-dioxane solution (2.0 mL) at room temperature, and the mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated, and ethyl acetate / n-hexane was added to the concentrated residue at room temperature. The precipitated solid was collected by filtration and dried under reduced pressure at room temperature, whereby 2-chloro-4- (thiophen-3-yl) -6,7,8,9-tetrahydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine hydrochloride (hereinafter, the compound of Example 5) was obtained as a yellow-brown solid (15 mg, yield 68%).
¹ H NMR (400 MHz, DMSO-d ₆ ) δ: 12.1 (1H, s), 9.22 (2H, brs), 7.85-7.37 (1H, m), 7.79-7.77 (1H, m), 7.38-7.37 ( 1H, m), 7.10 (1H, s), 3.95 (2H, brs), 3.50-3.47 (2H, m), 3.06 (2H, t, J = 5.2 Hz).
ESI-MS m / z 290 [M + H] ⁺ .

ｔｅｒｔ−ブチル ２−クロロ−４−（チオフェン−３−イル）−７，８−ジヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン−６（９Ｈ）−カルボキシレート（５０ｍｇ、０．１３ｍｍｏｌ）のトルエン（１０ｍＬ）溶液に、ｎ−プロピルボロン酸（１７ｍｇ、０．１９ｍｍｏｌ）、酢酸パラジウム（３．０ｍｇ、０．０１２ｍｍｏｌ）、２規定炭酸ナトリウム水溶液（０．１０ｍＬ）、トリシクロヘキシルホスフィン（５．５ｍｇ、０．０１９ｍｍｏｌ）を室温で加え、マイクロウエーブ照射下、１２０℃で１時間撹拌した。反応混合物を水に注ぎ、酢酸エチル（２５ｍＬ×３）で抽出した。有機層をあわせ、硫酸マグネシウムで乾燥後、濃縮した。得られた濃縮物をシリカゲルカラムクロマトグラフィー（溶出液：酢酸エチル／ｎ−ヘキサン）にて精製することで、ｔｅｒｔ−ブチル ２−プロピル−４−（チオフェン−３−イル）−７，８−ジヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン−６（９Ｈ）−カルボキシレートを灰色がかった白色固体として得た（１５ｍｇ、収率２９％）。
ESI-MS m/z 398 [M+H]⁺. Example 6 2-propyl-4- (thiophen-3-yl) -6,7,8,9-tetrahydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine hydrochloride Synthesis:
[Step 1] tert-Butyl 2-propyl-4- (thiophen-3-yl) -7,8-dihydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine-6 (9H) -Synthesis of carboxylates:

tert-butyl 2-chloro-4- (thiophen-3-yl) -7,8-dihydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine-6 (9H) -carboxylate ( To a solution of 50 mg, 0.13 mmol) in toluene (10 mL), n-propylboronic acid (17 mg, 0.19 mmol), palladium acetate (3.0 mg, 0.012 mmol), 2N aqueous sodium carbonate solution (0.10 mL), Tricyclohexylphosphine (5.5 mg, 0.019 mmol) was added at room temperature, and the mixture was stirred at 120 ° C. for 1 hour under microwave irradiation. The reaction mixture was poured into water and extracted with ethyl acetate (25 mL × 3). The organic layers were combined, dried over magnesium sulfate and concentrated. The resulting concentrate was purified by silica gel column chromatography (eluent: ethyl acetate / n-hexane) to give tert-butyl 2-propyl-4- (thiophen-3-yl) -7,8-dihydro. -5H-pyrrolo [2,3-b: 4,5-c '] dipyridine-6 (9H) -carboxylate was obtained as an off-white solid (15 mg, 29% yield).
ESI-MS m / z 398 [M + H] ⁺ .

ｔｅｒｔ−ブチル ２−プロピル−４−（チオフェン−３−イル）−７，８−ジヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン−６（９Ｈ）−カルボキシレート（１５ｍｇ、０．０３７ｍｍｏｌ）のジクロロメタン（２．０ｍＬ）溶液に、４規定塩化水素１，４−ジオキサン溶液（１．０ｍＬ）を室温で加え、室温で２時間撹拌した。反応混合物を濃縮し、濃縮残渣に酢酸エチル／ｎ−ヘキサンを室温で加えた。析出した固体をろ取し、室温で減圧乾燥することで、２−プロピル−４−（チオフェン−３−イル）−６，７，８，９−テトラヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン塩酸塩（以下、実施例６の化合物）を黄色固体として得た（７．０ｍｇ、収率６３％）。
¹H NMR (400 MHz, DMSO-d₆) δ: 11.6 (1H, s), 9.09 (2H, brs), 7.77-7.71 (2H, m), 7.33-7.32 (1H, m), 6.91 (1H, s), 3.94 (2H, brs), 3.49-3.38 (2H, m), 3.05-2.99 (2H, m), 2.76 (2H, t, J= 7.6 Hz), 1.77-1.70 (2H, m), 0.91 (3H, t, J = 7.6 Hz).
ESI-MS m/z 298 [M+H]⁺.[Step 2] Synthesis of 2-propyl-4- (thiophen-3-yl) -6,7,8,9-tetrahydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine hydrochloride :

tert-Butyl 2-propyl-4- (thiophen-3-yl) -7,8-dihydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine-6 (9H) -carboxylate ( To a solution of 15 mg, 0.037 mmol) in dichloromethane (2.0 mL) was added 4N hydrogen chloride 1,4-dioxane solution (1.0 mL) at room temperature, and the mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated, and ethyl acetate / n-hexane was added to the concentrated residue at room temperature. The precipitated solid was collected by filtration and dried under reduced pressure at room temperature, whereby 2-propyl-4- (thiophen-3-yl) -6,7,8,9-tetrahydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine hydrochloride (hereinafter, the compound of Example 6) was obtained as a yellow solid (7.0 mg, yield 63%).
¹ H NMR (400 MHz, DMSO-d ₆ ) δ: 11.6 (1H, s), 9.09 (2H, brs), 7.77-7.71 (2H, m), 7.33-7.32 (1H, m), 6.91 (1H, s), 3.94 (2H, brs), 3.49-3.38 (2H, m), 3.05-2.99 (2H, m), 2.76 (2H, t, J = 7.6 Hz), 1.77-1.70 (2H, m), 0.91 (3H, t, J = 7.6 Hz).
ESI-MS m / z 298 [M + H] ⁺ .

ｔｅｒｔ−ブチル ２−ブロモ−９−（（２−（トリメチルシリル）エトキシ）メチル）−７，８−ジヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン−６（９Ｈ）−カルボキシレート（１３０ｍｇ、０．２６ｍｍｏｌ）のＤＭＡ（３．０ｍＬ）溶液に、ナトリウム プロパン−２−チオレート（７９ｍｇ、０．８０ｍｍｏｌ）を室温で加え、８５℃で１６時間撹拌した。反応混合物に氷水を加え、酢酸エチル（２５ｍＬ×２）で抽出した。有機層をあわせ、硫酸ナトリウムで乾燥後、濃縮した。得られた濃縮物をシリカゲルカラムクロマトグラフィー（溶出液：メタノール／ジクロロメタン）にて精製することで、ｔｅｒｔ−ブチル ２−（イソプロピルチオ）−９−（（２−（トリメチルシリル）エトキシ）メチル）−７，８−ジヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン−６（９Ｈ）−カルボキシレートを灰色固体として得た（９８ｍｇ、収率８３％）。
ESI-MS m/z 478 [M+H]⁺.Reference Example 1 Synthesis of 2- (isopropylthio) -6,7,8,9-tetrahydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine hydrochloride:
[Step 1] tert-Butyl 2- (isopropylthio) -9-((2- (trimethylsilyl) ethoxy) methyl) -7,8-dihydro-5H-pyrrolo [2,3-b: 4,5-c ′ Synthesis of dipyridine-6 (9H) -carboxylate:

tert-Butyl 2-bromo-9-((2- (trimethylsilyl) ethoxy) methyl) -7,8-dihydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine-6 (9H) -Sodium propane-2-thiolate (79 mg, 0.80 mmol) was added to a solution of carboxylate (130 mg, 0.26 mmol) in DMA (3.0 mL) at room temperature, and the mixture was stirred at 85 ° C for 16 hours. Ice water was added to the reaction mixture, and the mixture was extracted with ethyl acetate (25 mL × 2). The organic layers were combined, dried over sodium sulfate, and concentrated. The obtained concentrate was purified by silica gel column chromatography (eluent: methanol / dichloromethane) to give tert-butyl 2- (isopropylthio) -9-((2- (trimethylsilyl) ethoxy) methyl) -7. , 8-dihydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine-6 (9H) -carboxylate was obtained as a gray solid (98 mg, 83% yield).
ESI-MS m / z 478 [M + H] ⁺ .

ｔｅｒｔ−ブチル ２−（イソプロピルチオ）−９−（（２−（トリメチルシリル）エトキシ）メチル）−７，８−ジヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン−６（９Ｈ）−カルボキシレート（９０ｍｇ、０．１８ｍｍｏｌ）のＴＨＦ（３．０ｍＬ）溶液に、ＴＢＡＦ（２ｍｏｌ／Ｌ ＴＨＦ溶液、４．０ｍＬ）を室温で加え、６０℃で１６時間撹拌した。反応混合物に水（２．０ｍＬ）を室温で加え、酢酸エチル（５．０ｍＬ×２）で抽出した。有機層をあわせ、硫酸ナトリウムで乾燥後、濃縮した。得られた濃縮物をシリカゲルカラムクロマトグラフィー（溶出液：メタノール／ジクロロメタン）にて精製することで、ｔｅｒｔ−ブチル ２−（イソプロピルチオ）−７，８−ジヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン−６（９Ｈ）−カルボキシレートを灰色固体として得た（５３ｍｇ、収率８０％）。
ESI-MS m/z 348 [M+H]⁺.[Step 2] Synthesis of tert-butyl 2- (isopropylthio) -7,8-dihydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine-6 (9H) -carboxylate:

tert-Butyl 2- (isopropylthio) -9-((2- (trimethylsilyl) ethoxy) methyl) -7,8-dihydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine-6 TBAF (2 mol / L THF solution, 4.0 mL) was added to a THF (3.0 mL) solution of (9H) -carboxylate (90 mg, 0.18 mmol) at room temperature, and the mixture was stirred at 60 ° C. for 16 hours. Water (2.0 mL) was added to the reaction mixture at room temperature, and the mixture was extracted with ethyl acetate (5.0 mL × 2). The organic layers were combined, dried over sodium sulfate, and concentrated. The obtained concentrate was purified by silica gel column chromatography (eluent: methanol / dichloromethane) to give tert-butyl 2- (isopropylthio) -7,8-dihydro-5H-pyrrolo [2,3-b. : 4,5-c ′] dipyridine-6 (9H) -carboxylate was obtained as a gray solid (53 mg, 80% yield).
ESI-MS m / z 348 [M + H] ⁺ .

ｔｅｒｔ−ブチル ２−（イソプロピルチオ）−７，８−ジヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン−６（９Ｈ）−カルボキシレート（５３ｍｇ、０．１５ｍｍｏｌ）に、４規定塩化水素１，４−ジオキサン溶液（３．０ｍＬ）を室温で加え、室温で２時間撹拌した。析出した固体をろ取し、酢酸エチル／ｎ−ヘキサン＝１／１で洗浄後、室温で減圧乾燥することで、２−（イソプロピルチオ）−６，７，８，９−テトラヒドロ−５Ｈ−ピロロ［２，３−ｂ：４，５−ｃ’］ジピリジン塩酸塩（以下、参考例１の化合物）を灰色がかった白色固体として得た（４３ｍｇ、収率９７％）。
¹H NMR (400 MHz, DMSO-d₆) δ: 11.68 (1H, brs), 9.68 (2H, brs), 7.77 (1H, d, J = 8.0 Hz), 6.94 (1H, d, J = 8.0 Hz), 4.26-4.20 (2H, m), 3.96-3.88 (1H, m), 3.47-3.40 (2H, m), 3.03-2.99 (2H, m), 1.33 (6H, d, J = 8.0 Hz).
ESI-MS m/z 248 [M+H]⁺.[Step 3] Synthesis of 2- (isopropylthio) -6,7,8,9-tetrahydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine hydrochloride:

To tert-butyl 2- (isopropylthio) -7,8-dihydro-5H-pyrrolo [2,3-b: 4,5-c ′] dipyridine-6 (9H) -carboxylate (53 mg, 0.15 mmol) A 4N hydrogen chloride 1,4-dioxane solution (3.0 mL) was added at room temperature, and the mixture was stirred at room temperature for 2 hours. The precipitated solid was collected by filtration, washed with ethyl acetate / n-hexane = 1/1, and then dried under reduced pressure at room temperature to give 2- (isopropylthio) -6,7,8,9-tetrahydro-5H-pyrrolo. [2,3-b: 4,5-c ′] dipyridine hydrochloride (hereinafter referred to as the compound of Reference Example 1) was obtained as an off-white solid (43 mg, yield 97%).
¹ H NMR (400 MHz, DMSO-d ₆ ) δ: 11.68 (1H, brs), 9.68 (2H, brs), 7.77 (1H, d, J = 8.0 Hz), 6.94 (1H, d, J = 8.0 Hz ), 4.26-4.20 (2H, m), 3.96-3.88 (1H, m), 3.47-3.40 (2H, m), 3.03-2.99 (2H, m), 1.33 (6H, d, J = 8.0 Hz).
ESI-MS m / z 248 [M + H] ⁺ .

実施例１と同様の手順により、実施例１のステップ１に記載のｔｅｒｔ−ブチル ４−オキソピペリジン−１−カルボキシレートの代わりにｔｅｒｔ−ブチル ４−オキソアゼパン−１−カルボキシレートを用いることで、２−（メチルチオ）−５，６，７，８，９，１０−ヘキサヒドロピリド［３’，２’：４，５］ピロロ［２，３−ｄ］アゼピン塩酸塩（以下、参考例２の化合物）を得た。
¹H NMR (400 MHz, DMSO-d₆) δ: 11.50 (1H, brs), 8.91 (2H, brs), 7.74 (1H, d, J = 8.2 Hz), 6.93 (1H, d, J = 8.2 Hz), 3.38-3.28 (4H, m), 3.10 (2H, t, J = 6.0 Hz), 3.01 (2H, t, J = 6.0 Hz), 2.52 (3H, s).
ESI-MS m/z 234 [M+H]⁺.Reference Example 2 2- (methylthio) -5,6,7,8,9,10-hexahydropyrido [3 ′, 2 ′: 4,5] pyrrolo [2,3-d] azepine hydrochloride Synthesis:

By using the same procedure as in Example 1, substituting tert-butyl 4-oxoazepane-1-carboxylate for tert-butyl 4-oxopiperidine-1-carboxylate described in Step 1 of Example 1, -(Methylthio) -5,6,7,8,9,10-hexahydropyrido [3 ', 2': 4,5] pyrrolo [2,3-d] azepine hydrochloride (hereinafter referred to as Reference Example 2) Compound) was obtained.
¹ H NMR (400 MHz, DMSO-d ₆ ) δ: 11.50 (1H, brs), 8.91 (2H, brs), 7.74 (1H, d, J = 8.2 Hz), 6.93 (1H, d, J = 8.2 Hz ), 3.38-3.28 (4H, m), 3.10 (2H, t, J = 6.0 Hz), 3.01 (2H, t, J = 6.0 Hz), 2.52 (3H, s).
ESI-MS m / z 234 [M + H] ⁺ .

Example 7 Human 5-HT2C receptor agonist activity:
Using cells stably expressing human 5-HT2C receptor (hereinafter, human 5-HT2C receptor-expressing cells), the 6-aza-tetrahydro-γ-carboline derivative represented by the above general formula (I) or a pharmacy thereof The human 5-HT2C receptor agonistic activity of the pharmaceutically acceptable acid addition salt was evaluated.

  By excising the human 5-HT2C receptor gene site of the plasmid (GenBank accession number: AB5288036.1) into which the human 5-HT2C receptor gene has been inserted and cloning it into the mammalian cell expression vector pCI-neo (Promega) The prepared pCI-neo-HTR2C was introduced into HEK-293 cells (American Type Culture Collection). A single clone was obtained by limiting dilution from cells into which the expression vector pCI-neo-HTR2C had been introduced, and human 5-HT2C receptor-expressing cells were prepared.

Human 5-HT2C receptor-expressing cells are Dulbecco's Modified Eagle containing 10% fetal bovine serum (dialyzed), 1% non-essential amino acid for MEM, 100 U / mL penicillin, 100 μg / mL streptomycin and 500 μg / mL Geneticin. The culture medium was maintained in a 5% CO ₂ incubator at 37 ° C. using 's Medium as a culture solution.

Human 5-HT2C receptor-expressing cells are suspended in the above culture medium (but not Geneticin) and seeded at 8 × 10 ⁴ cells in each well of a 96-well black plate (Corning) at 37 ° C., 5% Cultivated overnight in CO ₂ and used for the following evaluation.

  Evaluation of human 5-HT2C receptor agonistic activity was performed by measuring an increase in intracellular calcium concentration due to activation of human 5-HT2C receptor.

  For the measurement of intracellular calcium concentration, FLIPR (registered trademark) Calcium 5 Assay Kit (Molecular Devices) was used. As the assay buffer, Hank's balanced salt solution containing 20 mmol / L HEPES (pH 7.3) added with 2.5 mmol / L probenecid (SIGMA) was used.

The medium of the plate seeded with human 5-HT2C receptor-expressing cells was removed, and FLIPR (registered trademark) Calcium5 Assay Reagent (included in the Kit) 150 μL / well dissolved in the assay buffer according to the instructions attached to the Kit In addition, the cells were incubated at 37 ° C. and 5% CO ₂ for 60 minutes under light shielding, and further allowed to stand at room temperature for 15 minutes under light shielding. Using FLIPR (registered trademark) TETRA (Molecular Devices), 50 μL / well of a test compound was automatically added, and the fluorescence intensity was measured at an excitation wavelength of 470-495 nm and a fluorescence wavelength of 515-575 nm for 5 minutes.

Evaluation of each test compound was carried out by triplicate at each concentration at a common ratio of 3. The fluorescence intensity at the time of assay buffer addition was defined as 0% response value, and the maximum fluorescence intensity when serotonin (Nacalai Tesque), a human 5-HT2C receptor agonist, was added instead of the test compound was defined as 100% response value. The reaction rate (%) at each concentration of the test compound was determined. The EC ₅₀ value of each test compound (concentration showing 50% response to 100% response value of serotonin) was determined by linear regression using the reaction rate (%) at each concentration of each test compound. Each test compound and serotonin were dissolved in DMSO and then diluted with an assay buffer.

The EC ₅₀ values for each test compound are shown in Table 2. As is apparent from the results in Table 2, the 6-aza-tetrahydro-γ-carboline derivative represented by the above general formula (I) or a pharmaceutically acceptable acid addition salt thereof is the same as the compounds of Reference Examples 1 and 2. In comparison, all were shown to have potent human 5-HT2C receptor agonist activity.

(Example 8) Effect on pressure at leaking rat under anesthesia:
The 6-aza-tetrahydro-γ-carboline derivative represented by the above general formula (I) or a pharmaceutically acceptable acid thereof by using a pressure measurement method in anesthetized rats that can evaluate the effect on stress urinary incontinence The effect of the addition salt was evaluated.

  SD female rats (Nippon Charles River) weighing 190-340 g were anesthetized with isoflurane (Mylan Pharmaceutical), and the spinal cord was cut at the T8-T9 level for the purpose of eliminating the micturition reflex. Next, the lower abdomen was incised, the ureters on both sides were ligated, and the kidney side was cut. In addition, two catheters (PE-90 and PE-50; Becton & Dickinson) were placed in the bladder. Immediately before waking up from anesthesia, urethane (SIGMA) was additionally anesthetized in the abdominal cavity, and then the abdominal muscles and skin incisions were sutured. The PE-50 type intravesical catheter was connected to a pressure transducer, and the PE-90 type catheter was connected to a 60 mL syringe filled with physiological saline. The transducer signal was sent to a computer via an amplifier (blood pressure amplification unit AP-641G; Nihon Kohden) and a data analyzer (PowerLab; AD Instruments Inc.), recorded on a hard disk, and analyzed. The physiological saline was injected into the bladder by vertically raising a 60 mL syringe filled with physiological saline, and the maximum intravesical pressure (leakage pressure) when physiological saline leaked from the urethral orifice was measured. After confirming that the physiological saline solution leaked from the urethral orifice, the bladder infusion of the physiological saline solution was stopped, and the physiological saline solution retained in the bladder was discharged.

  The bladder was infused and excreted with physiological saline, and the test compound was administered into the tail vein immediately after a stable leak pressure was obtained three times. Saline solution was injected into the urinary bladder 5 minutes after administration of the test compound, and the leakage pressure was measured. The rate of change (%) in leakage pressure after administration with respect to the average value of leakage pressure three times immediately before test compound administration was used as the evaluation value.

  The compound of Example 1 was used as the 6-aza-tetrahydro-γ-carboline derivative represented by the above general formula (I) or a pharmaceutically acceptable acid addition salt thereof. The compound of Example 1 was dissolved in physiological saline and administered at 0.3 mg / kg (volume 0.5 mL / kg). As a control, duloxetine, a therapeutic agent for stress urinary incontinence, was used. Duloxetine (Kemprotec) was dissolved in distilled water and administered at 1 mg / kg (volume 0.5 mL / kg). In addition, as a control for the compound of Example 1 and duloxetine, a group in which the same volume of each solvent was administered was provided.

  The results are shown in FIG. The vertical axis shows the rate of change (%) in the leaking pressure after administration of the test compound (mean ± standard error; N = 6 to 12). In FIG. 1, * marks indicate statistically significant (p <0.05, t test) in comparison with each control group.

  The compound of Example 1 (0.3 mg / kg) and duloxetine (1 mg / kg) increased the leakage pressure statistically significantly compared to the respective control groups.

  From this result, the 6-aza-tetrahydro-γ-carboline derivative represented by the above general formula (I) or a pharmaceutically acceptable acid addition salt thereof is treated or prevented for stress urinary incontinence in the same manner as duloxetine. It was shown to be effective.

  Since the 6-aza-tetrahydro-γ-carboline derivative represented by the above general formula (I) or a pharmaceutically acceptable acid addition salt thereof has a potent 5-HT2C receptor agonist activity, It can be used as a therapeutic or preventive agent for pressure urinary incontinence.

Claims (7)

A 6-aza-tetrahydro-γ-carboline derivative represented by the following general formula (I) or a pharmaceutically acceptable acid addition salt thereof.

[Wherein, R ¹ represents an alkyl group having 1 to 6 carbon atoms, an alkylthio group having 1 or 2 carbon atoms, or a halogen atom, and R ² represents a hydrogen atom or an alkyloxy group having 1 to 3 carbon atoms. Or a heteroaryl group having 5 or 6 ring atoms, R ³ represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms, and R ¹ is an alkyl group or halogen atom having 1 to 6 carbon atoms. In this case, R ² is an alkyloxy group having 1 to 3 carbon atoms or a heteroaryl group having 5 or 6 ring atoms. ] R ¹ is an alkyl group having 1 to 6 carbon atoms, an alkylthio group having 1 or 2 carbon atoms, or a chlorine atom, and R ² is a hydrogen atom, an alkyloxy group having 1 to 3 carbon atoms, or 3- The 6-aza-tetrahydro-γ-carboline derivative or a pharmaceutically acceptable acid addition salt thereof according to claim 1, which is a thienyl group and R ³ is a hydrogen atom. The 6-aza-tetrahydro-γ-carboline derivative according to claim 1, wherein R ¹ is an n-propyl group, R ² is a methoxy group or a 3-thienyl group, and R ³ is a hydrogen atom. A pharmaceutically acceptable acid addition salt thereof. The 6-aza-tetrahydro-γ-carboline derivative according to claim 1, wherein R ¹ is a methylthio group, R ² is a hydrogen atom or a methoxy group, and R ³ is a hydrogen atom, or a pharmaceutically acceptable salt thereof. Acceptable acid addition salt.   A medicament comprising the 6-aza-tetrahydro-γ-carboline derivative according to any one of claims 1 to 4 or a pharmaceutically acceptable acid addition salt thereof as an active ingredient.   A 5-HT2C receptor agonist comprising the 6-aza-tetrahydro-γ-carboline derivative according to any one of claims 1 to 4 or a pharmaceutically acceptable acid addition salt thereof as an active ingredient.   The therapeutic agent or preventive agent of stress urinary incontinence which contains the 6-aza-tetrahydro-gamma-carboline derivative or its pharmaceutically acceptable acid addition salt as described in any one of Claims 1-4 as an active ingredient. .

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