HK1217171B - V1a antagonists to treat phase shift sleep disorders - Google Patents

Description

Via antagonists for the treatment of phase-shift sleep disorders

Overview

The present invention relates to novel medical uses of certain chemical compounds and pharmaceutical compositions containing the same. The present invention relates to compounds that are Via antagonists for treating phase-shift sleep disorders, particularly jet lag. In another aspect, the present invention relates to pharmaceutical compositions for treating phase-shift sleep disorders, comprising a compound according to the present invention and a pharmaceutically acceptable carrier.

Background Art

The suprachiasmatic nucleus (SCN) is the body's intrinsic clock that regulates circadian rhythms and is known to be rich in vasopressin neurons (Kalsbeek et al. 2010). ¹ Vasopressin is produced and released in a 24-hour rhythm (Schwartz et al. 1983). ² A primary regulatory role for vasopressin in circadian rhythms has not been demonstrated by prior art. Brattleboro rats, a rat strain congenitally deficient in vasopressin due to a point mutation, have no apparent defects in their circadian rhythms (Groblewski et al. 1981). ³ Direct injection of vasopressin into the hamster SCN has no effect on circadian phase shifts (Albers et al. 1984). ⁴ Instead, vasopressin may regulate the circadian clock in more subtle ways. Vasopressin-deficient Brattleboro rats did not respond to the 6-h phase advance of the light-dark period but remained synchronized to food delivery (Murphy et al. 1998) ⁵ , in contrast to normal rats, which responded to both the novel light-dark rhythm and food delivery.

More specifically, studies of V1a knockout (KO) mice have shown that the vasopressin V1a receptor regulates circadian behavior. V1a KO mice behave normally under a normal 12h-12h light-dark cycle, but in the absence of light (dark-dark conditions), these mice show a gradual loss of circadian rhythmicity, characterized by a prolonged active phase. However, they still respond normally to phase shifts caused by brief light exposure during the dark phase (Li et al. 2009) ^.

We surprisingly found that administration of a single dose of a small molecule Via antagonist to normal mice before a 6h phase shift (6h advance of light phase) resulted in significantly faster re-entrainment to the new light-dark cycle compared to vehicle-treated mice. More specifically, a selective Via antagonist.

This finding is contrary to the teaching of the prior art, in which treatment with melatonin to accelerate readaptation after a phase shift was effective only after 3 days of daily administration following a phase advance (Dubocovich et al. 2005) ⁷ .

Poor sleep can lead to a variety of health disturbances, including anxiety, depression, irritability, and impaired social and psychomotor coordination.

WO 2013/176220 ⁸ describes circadian rhythm regulators, which include inhibitors capable of inhibiting vasopressin receptors V1a and V1b.

Via antagonists have been described in the prior art, for example in EP 0382185, EP 0526348, WO 94/01113, WO 94/18975, WO 95/06035, WO 96/22292, WO 96/22293, WO 97/49707, WO 97/49708, WO 98/24430, WO 99/37637, WO 99/44613, WO 99/55340, WO 99/65525, WO 00/029405, WO 00/066117, WO 01/058880, WO 01/066109, WO 02/002531, WO 02/044179, WO 02/055514, WO 03/031407, WO 03/037901, WO 03/042181, WO 04/074291, WO 04/108138, WO 2005/039565, WO 2005/063754, WO 2006/020491, WO 2006/021213, WO 2006/051851, WO 2006/058705, WO 2006/102283, WO 2006/102308, WO 2007/006688, WO 2007/009906, WO 2007/014851, WO 2007/039438, WO 2007/077122, WO 2007/109615, WO 2008/068159, WO2008/068183, WO 2008/068184, WO 2008/068185, WO 2008/077810, WO 2008/077811, WO2008/080842, WO 2008/080844, WO 2008/084005, WO 2010/057795, WO 2010/060836, WO2011/120877, WO 2011/128265, WO 2011/131596,WO 2011/134877, WO 2011/141396, WO2012/003436, WO 2012/042534, WO 2013/045373, WO 2013/050334 ⁹ etc.

Detailed Description of the Invention

The following definitions of the general terms used in this specification apply regardless of whether the terms appear alone or in combination with other groups.

The term "phase shift sleep disorders" encompasses conditions that are classified as disturbances of the circadian rhythm, ie the approximately 24-hour cycle produced by an organism, such as a human.

Phase shift sleep disorders include, but are not limited to, temporary disorders such as jet lag or altered sleep schedules due to work, social responsibilities, or illness, as well as chronic disorders such as delayed sleep-phase syndrome (DSPS), delayed sleep-phase type (DSPT), advanced sleep-phase syndrome (ASPS), and irregular sleep-wake cycles.

The term "V1a antagonist" refers to an antagonist of the V1a vasopressin receptor subtype. Examples include OPC-21268, OPC-31260, OPC-41061, VPA-985, SR-49059, SR-121463, VP-343, FR-161282, CI-1025, SRX-251, SRX-246, JNJ-17158063, FE-202158, VT-913, LY307174, Org-52186, PF-184563, YM-218, 1-({(2R,3S)-5-chloro-3-(2-chlorophenyl)-1-[(3,4-dimethoxyphenyl)sulfonyl]-3-hydroxy-2 ... , 3-dihydro-1H-indol-2-yl}carbonyl)-L-prolinamide, [6-chloro-1-[2-(dimethylamino)ethyl]indol-3-yl]-spiro[1H-isobenzofuran-3,4′-piperidin]-1′-yl-methanone, 8-chloro-5-methyl-1-(3,4,5,6-tetrahydro-2H-[1,2′]bipyridin-4-yl)-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azulene, trans-8-chloro-5-methyl-1-(4-pyridin-2-yloxycyclohexyl)-4,6-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine, and the like. The term “selective Via antagonist” refers to selectivity for Via over Vib receptor subtypes. An example is trans-8-chloro-5-methyl-1-(4-pyridin-2-yloxycyclohexyl)-4,6-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

As used herein, the term "alkyl", alone or in combination with other groups, refers to a saturated (i.e., aliphatic) hydrocarbon group containing a straight or branched carbon chain. If not further specified, "alkyl" refers to a group having 1 to 12 carbon atoms, such as "Ci _-i2 -alkyl". "Ci _-4 -alkyl" refers to an alkyl group having 1 to 4 carbon atoms and "Ci _-7 -alkyl" refers to an alkyl group having 1 to 7 carbon atoms. Examples of "alkyl" are methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, and the like. Preferred are methyl and isopropyl.

The term "alkoxy," alone or in combination with other groups, refers to the group -O-R', where R' is an alkyl group as defined above. "Ci _-i2 -alkoxy" refers to an alkoxy group having 1 to 12 carbon atoms, "Ci _-4 -alkoxy" refers to an alkoxy group having 1 to 4 carbon atoms, and "Ci _-7 -alkoxy" refers to an alkoxy group having 1 to 7 carbon atoms. Examples of "alkoxy" are methoxy, ethoxy, propoxy, tert-butoxy, and the like. Preferred is methoxy.

The term "aromatic" denotes the presence of an electron sextet in the ring according to Hückel's rule.

The term "cyano" refers to the group -CN.

The term "hydroxy" refers to the group -OH.

The term "halo" or "halogen" refers to chlorine, iodine, fluorine and bromine. Chlorine and fluorine are preferred.

The terms "halo-Ci _-n -alkyl" and "Ci _-n -haloalkyl", alone or in combination with other groups, mean a Ci _-n -alkyl group as defined above, having 1 to n carbon atoms as defined in the specification, wherein at least one of the hydrogen atoms of the alkyl group is replaced by a halogen atom (preferably fluorine or chlorine, most preferably fluorine). Examples of halo-Ci _-n -alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, isobutyl, sec-butyl, tert-butyl, pentyl or n-hexyl substituted by one or more Cl, F, Br or I atoms, especially one _, two or three fluorine or chlorine, and those groups specifically illustrated by the examples herein below. Preferred halo- _Ci-n -alkyl groups include difluoro- or trifluoro-methyl or -ethyl, and _-CF3 , -CH( _CH3 ) _CH2CF3 , -CH( _CH3 ) _CH2F .

The term "heterocycloalkyl" as defined herein, alone or in combination with other groups, refers to a monovalent 3 to 7 membered or 4 to 7 membered saturated ring containing one or two heteroatoms selected from N, O or S. The term "3 to 7 membered heterocycloalkyl" as defined herein, alone or in combination with other groups, refers to a monovalent 3 to 7 membered ring containing one or two heteroatoms selected from N, O or S. The term "4-7 membered heterocycloalkyl" as defined herein, alone or in combination with other groups, refers to a 4 to 7 membered saturated ring containing one or two heteroatoms selected from N, O or S. Examples of heterocycloalkyl moieties are oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, pyrrolidinyl, morpholinyl, thiomorpholinyl, piperidinyl, or piperazinyl. Preferred heterocycloalkyls are oxetanyl and tetrahydrofuranyl. Heterocycloalkyls are optionally substituted as described herein.

The term "heteroaryl", alone or in combination with other groups, refers to a monovalent aromatic 5- or 6-membered monocyclic ring ("5- or 6-membered heteroaryl") containing one or two ring heteroatoms selected from N, O, or S, with the remaining ring atoms being C. 6-membered heteroaryl is preferred. Examples of heteroaryl moieties include, but are not limited to, pyridinyl, pyrimidinyl, or pyrazinyl. Pyridinyl is preferred.

The term "aryl," alone or in combination with other groups, means a monovalent cyclic aromatic hydrocarbon moiety consisting of a monocyclic or bicyclic aromatic ring. Particular aryl groups are phenyl or naphthyl. Aryl groups can be unsubstituted or substituted as described herein.

The terms "cycloalkyl" and " _C3-7 -cycloalkyl", alone or in combination with other groups, refer to a 3- to 7-membered carbon ring, for example cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

The term "oxo" when referring to a substituent on a heterocycloalkyl group means that an oxygen atom is attached to the heterocycloalkyl ring. Thus, an "oxo" group may replace two hydrogen atoms on a carbon atom, or it may simply be attached to sulfur so that the sulfur is present in an oxidized form, i.e., with one or two oxygen atoms, such as the group _-SO2 .

When indicating the number of substituents, the term "one or more" means from one substituent to the maximum possible number of substitutions, i.e., from replacement of one hydrogen by a substituent to replacement of all hydrogens by a substituent. Thus, one, two or three substituents are preferred. Even more preferred are one or two substituents or one substituent.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below.

The terms "pharmaceutically acceptable carrier" and "pharmaceutically acceptable auxiliary substance" refer to carriers and auxiliary substances such as diluents or excipients that are compatible with the other ingredients of the formulation. Such formulations are formulations in pharmaceutical form.

The term "prodrug" refers to a structural derivative of a drug that must be chemically converted into the drug in vivo in order to exert its pharmacological or therapeutic effect (see Patrick ¹⁰ or Ganellin et al. ¹¹ ).

The term "pharmaceutical composition" includes a product comprising specified ingredients in predetermined amounts or proportions, as well as any product that results, directly or indirectly, from combining the specified ingredients in the specified amounts. In particular, it includes a product comprising one or more active ingredients and, optionally, a carrier including an inert ingredient, as well as any product that results, directly or indirectly, from combining, complexing, or aggregating any two or more of the ingredients, or from decomposing one or more of the ingredients, or from some other type of reaction or interaction of one or more of the ingredients.

"Therapeutically effective amount" means the amount of a compound that, when administered to a patient for treating a disease state, is sufficient to effect such treatment for the disease state. The "therapeutically effective amount" will vary depending on the compound, the disease state being treated, the severity or illness being treated, the age and relative health of the patient, the route and form of administration, the judgment of the attending physician or veterinarian, and other factors.

The terms "as defined herein" and "as described herein" when referring to a variable incorporate by reference the broad definition of that variable as well as the particular, more particular and most particular definitions, if any.

The terms "treating," "contacting," and "reacting" when referring to a chemical reaction mean adding or mixing two or more reagents under appropriate conditions to produce a desired and/or intended product. It should be understood that a reaction that produces a desired and/or intended product may not necessarily result directly from the combination of the two initially added reagents, i.e., one or more intermediates may be present that are produced in the mixture and ultimately lead to the formation of the desired and/or intended product. Treatment includes prophylactic treatment as well as immediate relief of symptoms.

The term "aromatic" refers to the conventional concept of aromaticity as defined in the literature, in particular IUPAC ¹² .

The term "pharmaceutically acceptable excipient" refers to any ingredient that is not therapeutically active and non-toxic such as a disintegrant, binder, filler, solvent, buffer, tonicity agent, stabilizer, antioxidant, surfactant or lubricant used in formulating a pharmaceutical product.

The corresponding pharmaceutically acceptable salts of acids can be obtained by standard methods known to those skilled in the art, for example, by dissolving the compound of formula I in a suitable solvent such as dioxane or THF, and adding an appropriate amount of the corresponding acid. The product can usually be separated by filtration or chromatography. The conversion of a compound of formula (I) to a pharmaceutically acceptable salt using a base can be carried out by treating such a compound with such a base. A possible method for forming such a salt is, for example, by adding 1/n equivalents of an alkaline salt such as, for example, M(OH) _n (wherein M=metal or ammonium cation and n=number of hydroxide anions) to a solution of the compound in a suitable solvent (e.g., ethanol, ethanol-water mixture, tetrahydrofuran-water mixture), and removing the solvent by evaporation or lyophilization.

One embodiment of the present invention relates to the use of a Via antagonist for the treatment and/or prevention of phase shift sleep disorders.

One embodiment of the present invention relates to the use of a Via antagonist for the treatment of phase shift sleep disorders.

One embodiment of the present invention relates to the use of a Via antagonist for preventing phase shift sleep disorders.

One embodiment of the present invention relates to the use of a Via antagonist for the treatment and/or prevention of a phase shift sleep disorder, wherein the phase shift sleep disorder is jet lag.

One embodiment of the invention relates to the use of a Via antagonist, wherein the Via antagonist is a compound of formula I, for the treatment and/or prevention of phase shift sleep disorders.

in

R ¹ is aryl or heteroaryl, which is unsubstituted or substituted with one or more substituents independently selected from A,

^R2 is

H,

● C _1-12 -alkyl, which is unsubstituted or substituted by one or more OH, halogen, cyano or C _1-12 -alkoxy,

-(CH ₂ ) _q -R ^a , wherein R ^a is phenyl or a 5-membered or 6-membered heteroaryl, each of which is unsubstituted or substituted with one or more substituents independently selected from A,

●-(CH ₂ ) _r NR ⁱ R ⁱⁱ ，

-C(O)-C _1-12 -alkyl, where C _1-12 -alkyl is unsubstituted or substituted by one or more OH, halogen, cyano or C _1-12 -alkoxy groups,

●-C(O)(CH ₂ ) _q OC(O)-C _1-12 -alkyl,

●-C(O)(CH ₂ ) _q NR ⁱ R ⁱⁱ ,

-C(O)OC _1-12 -alkyl, wherein alkyl is unsubstituted or substituted by one or more OH, halogen, cyano or C _1-12 -alkoxy groups,

-S(O) ₂ -C _1-12 -alkyl, or

●-S(O) ₂ NR ⁱ R ⁱⁱ ，

R ⁱ and R ⁱⁱ are each independently H, C _1-12 -alkyl, or together with the nitrogen to which they are attached form a 3- to 7-membered heterocycloalkyl containing one or two heteroatoms selected from N, O or S, said heterocycloalkyl being unsubstituted or substituted with one or more substituents independently selected from B,

q is 1, 2, 3, or 4,

r is 2, 3, or 4,

A is halogen, cyano, OH, C _1-7 -alkyl, halo-C _1-7 -alkyl or C _1-7 -alkoxy, halo-C _1-7 -alkoxy or hydroxy-C _1-7 -alkyl,

B is oxo, halogen, OH, C _1-7 -alkyl or C _1-7 -alkoxy,

^R3 is Cl or F,

or a pharmaceutically acceptable salt thereof.

One embodiment of the present invention relates to the use of a selective Via antagonist for the treatment and/or prevention of phase shift sleep disorders.

One embodiment of the present invention relates to the use of a selective Via antagonist for the treatment of phase shift sleep disorders.

One embodiment of the present invention relates to the use of a selective Via antagonist for preventing phase shift sleep disorders.

One embodiment of the present invention relates to the use of a selective Via antagonist for the treatment and/or prevention of a phase shift sleep disorder, wherein the phase shift sleep disorder is jet lag.

One embodiment of the invention relates to the use of a Via antagonist for the treatment and/or prevention of phase shift sleep disorders, wherein ^R1 is a monovalent cyclic aromatic hydrocarbon moiety consisting of a monocyclic or bicyclic aromatic ring; or a monovalent 5-membered or 6-membered aromatic monocyclic or 9-membered or 10-membered aromatic bicyclic ring containing one to four ring heteroatoms selected from N, O or S, the remaining ring atoms being C; each of which is unsubstituted or substituted by one or more substituents independently selected from A; and A is halogen, cyano, OH, _C1-7 -alkyl, halo- _C1-7 -alkyl, _C1-7 -alkoxy, halo- _C1-7 -alkoxy or hydroxy- _C1-7 -alkyl.

One embodiment of the present invention relates to the use of a Via antagonist for the treatment and/or prevention of phase shift sleep disorders, wherein the Via antagonist is a compound of formula I, wherein R ¹ is a monovalent cyclic aromatic hydrocarbon moiety consisting of a mono-aromatic ring.

One embodiment of the invention relates to the use of a Via antagonist for the treatment and/or prevention of phase shift sleep disorders, wherein said Via antagonist is a compound of formula I, wherein R ¹ is pyridinyl.

One embodiment of the present invention relates to the use of a Via antagonist for the treatment and/or prevention of phase shift sleep disorders, wherein the Via antagonist is a compound of formula I, wherein ^R2 is

H,

● C _1-12 -alkyl, which is unsubstituted or substituted by one or more OH groups,

-(CH ₂ ) _q -R ^a , wherein R ^a is phenyl or a 5-membered or 6-membered heteroaryl group and q is 1, 2, 3 or 4, preferably 1,

-C(O)-C _1-12 -alkyl,

-C(O)(CH ₂ ) _q NR ⁱ R ⁱⁱ , wherein R ⁱ and R ⁱⁱ are each independently H or C _1-12 -alkyl, preferably C _1-12 -alkyl, and q is 1, 2, 3 or 4, preferably 1,

-C(O)OC _1-12 -alkyl,

-S(O) ₂ -C _1-12 -alkyl, or

-S(O) ₂ NR ⁱ R ⁱⁱ , wherein R ⁱ and R ⁱⁱ are each independently H or C _1-12 -alkyl, preferably C _1-12 -alkyl.

One embodiment of the invention relates to the use of a Via antagonist for the treatment and/or prevention of phase shift sleep disorders, wherein said Via antagonist is a compound of formula I, wherein ^R2 is methyl.

One embodiment of the invention relates to the use of a Via antagonist for the treatment and/or prevention of phase shift sleep disorders, wherein said Via antagonist is a compound of formula I, wherein R ³ is Cl.

One embodiment of the present invention relates to the use of a Via antagonist for the treatment and/or prevention of phase shift sleep disorders, wherein the Via antagonist is a compound of formula I, wherein the compound is selected from the group consisting of:

trans-8-chloro-1-(4-phenoxy-cyclohexyl)-4H,6H-2,3,5,10b-tetraaza-benzoazulene-5-carboxylic acid tert-butyl ester,

trans-8-chloro-1-(4-phenoxy-cyclohexyl)-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoazulene hydrochloride,

trans-8-chloro-5-methyl-1-(4-phenoxy-cyclohexyl)-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azulene,

trans-1-[8-chloro-1-(4-phenoxy-cyclohexyl)-4H,6H-2,3,5,10b-tetraaza-benzoazulene-5-yl]-ethanone,

trans-8-chloro-5-methanesulfonyl-1-(4-phenoxy-cyclohexyl)-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoazulene,

trans-2-[8-chloro-1-(4-phenoxy-cyclohexyl)-4H,6H-2,3,5,10b-tetraaza-benzoazulene-5-yl]-ethanol,

trans-8-chloro-5-isopropyl-1-(4-phenoxy-cyclohexyl)-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoazulene,

trans-8-chloro-1-(4-phenoxy-cyclohexyl)-4H,6H-2,3,5,10b-tetraaza-benzoazulene-5-sulfonic acid dimethylamide,

trans-8-chloro-1-(4-phenoxy-cyclohexyl)-5-pyridin-2-ylmethyl-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoazulene,

trans-1-[8-chloro-1-(4-phenoxy-cyclohexyl)-4H,6H-2,3,5,10b-tetraaza-benzoazulene-5-yl]-2-dimethylamino-ethanone,

trans-8-Fluoro-1-(4-phenoxy-cyclohexyl)-4H,6H-2,3,5,10b-tetraaza-benzo[e]azulene-5-carboxylic acid tert-butyl ester,

trans-8-fluoro-5-methyl-1-(4-phenoxy-cyclohexyl)-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoazulene,

cis-8-Chloro-1-(4-phenoxy-cyclohexyl)-4H,6H-2,3,5,10b-tetraaza-benzoazulene-5-carboxylic acid tert-butyl ester,

cis-8-chloro-5-methyl-1-(4-phenoxy-cyclohexyl)-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoazulene,

trans-8-chloro-1-[4-(4-fluoro-phenoxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzoazulene-5-carboxylic acid tert-butyl ester,

trans-8-chloro-1-[4-(4-fluoro-phenoxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azulene hydrochloride,

trans-8-chloro-1-[4-(4-fluoro-phenoxy)-cyclohexyl]-5-methyl-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoazulene,

trans-8-chloro-1-[4-(4-cyano-phenoxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzo[e]azulene-5-carboxylic acid tert-butyl ester,

trans-4-[4-(8-chloro-5-methyl-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azulen-1-yl)-cyclohexyloxy]-benzonitrile,

trans-8-chloro-1-[4-(4-trifluoromethyl-phenoxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzo[e]azulene-5-carboxylic acid tert-butyl ester,

trans-8-chloro-1-[4-(3-chloro-phenoxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzoazulene-5-carboxylic acid tert-butyl ester,

trans-8-chloro-1-[4-(3-chloro-phenoxy)-cyclohexyl]-5-methyl-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoazulene,

trans-8-chloro-1-[4-(3-methoxy-phenoxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzoazulene-5-carboxylic acid tert-butyl ester,

trans-8-chloro-1-[4-(3-methoxy-phenoxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azulene hydrochloride,

trans-8-chloro-1-[4-(3-methoxy-phenoxy)-cyclohexyl]-5-methyl-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoazulene,

trans-3-[4-(8-chloro-5-methyl-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoazulen-1-yl)-cyclohexyloxy]-benzonitrile,

trans-8-chloro-1-(4-m-tolyloxy-cyclohexyl)-4H,6H-2,3,5,10b-tetraaza-benzo[e]azulene-5-carboxylic acid tert-butyl ester,

trans-8-chloro-1-(4-m-tolyloxy-cyclohexyl)-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azulene hydrochloride,

trans-8-chloro-5-methyl-1-(4-m-tolyloxy-cyclohexyl)-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azulene,

trans-1-[4-(3-tert-butyl-phenoxy)-cyclohexyl]-8-chloro-4H,6H-2,3,5,10b-tetraaza-benzo[e]azulene-5-carboxylic acid tert-butyl ester,

trans-8-chloro-1-[4-(2-fluoro-phenoxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzo[e]azulene-5-carboxylic acid tert-butyl ester,

trans-8-chloro-1-[4-(2-fluoro-phenoxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azulene hydrochloride,

trans-8-chloro-1-[4-(2-fluoro-phenoxy)-cyclohexyl]-5-methyl-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azulene,

trans-8-chloro-1-[4-(2-cyano-phenoxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzoazulene-5-carboxylic acid tert-butyl ester,

trans-2-[4-(8-chloro-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azulen-1-yl)-cyclohexyloxy]-benzonitrile hydrochloride,

trans-2-[4-(8-chloro-5-methyl-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoazulen-1-yl)-cyclohexyloxy]-benzonitrile,

trans-8-chloro-1-(4-o-tolyloxy-cyclohexyl)-4H,6H-2,3,5,10b-tetraaza-benzoazulene-5-carboxylic acid tert-butyl ester,

trans-8-chloro-5-methyl-1-(4-o-tolyloxy-cyclohexyl)-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoazulene,

trans-8-chloro-1-[4-(3,5-difluoro-phenoxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzoazulene-5-carboxylic acid tert-butyl ester,

trans-8-chloro-1-[4-(3,5-difluoro-phenoxy)-cyclohexyl]-5-methyl-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoazulene,

trans-8-chloro-1-[4-(naphthalen-2-yloxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzoazulene-5-carboxylic acid tert-butyl ester,

trans-8-chloro-5-methyl-1-[4-(naphthalen-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoazulene,

trans-8-chloro-1-[4-(pyridin-3-yloxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzoazulene-5-carboxylic acid tert-butyl ester,

trans-8-chloro-5-methyl-1-[4-(pyridin-3-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoazulene,

trans-8-chloro-1-[4-(5-chloro-pyridin-3-yloxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzoazulene-5-carboxylic acid tert-butyl ester,

trans-8-chloro-1-[4-(5-chloro-pyridin-3-yloxy)-cyclohexyl]-5-methyl-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoazulene,

trans-8-chloro-1-[4-(pyridin-2-yloxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzoazulene-5-carboxylic acid tert-butyl ester,

trans-8-chloro-1-[4-(pyridin-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoazulene hydrochloride,

trans-8-chloro-5-methyl-1-[4-(pyridin-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoazulene,

trans-8-chloro-1-[4-(6-chloro-pyridin-2-yloxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzo[e]azulene-5-carboxylic acid tert-butyl ester,

trans-8-chloro-1-[4-(6-chloro-pyridin-2-yloxy)-cyclohexyl]-5-methyl-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azulene,

trans-8-chloro-1-[4-(5-chloro-pyridin-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azulene hydrochloride,

trans-8-chloro-1-[4-(5-chloro-pyridin-2-yloxy)-cyclohexyl]-5-methyl-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azulene,

trans-8-chloro-1-[4-(pyrimidin-2-yloxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzo[e]azulene-5-carboxylic acid tert-butyl ester,

cis-8-chloro-1-[4-(pyrimidin-2-yloxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzo[e]azulene-5-carboxylic acid tert-butyl ester,

trans-8-chloro-5-methyl-1-[4-(pyrimidin-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azulene,

trans-8-chloro-1-[4-(pyrazin-2-yloxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzo[e]azulene-5-carboxylic acid tert-butyl ester,

trans-8-chloro-5-methyl-1-[4-(pyrazin-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azulene,

trans-8-chloro-1-[4-(pyrimidin-4-yloxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzo[e]azulene-5-carboxylic acid tert-butyl ester,

trans-8-chloro-5-methyl-1-[4-(pyrimidin-4-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azulene,

trans-8-chloro-1-[4-(pyridazin-3-yloxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzo[e]azulene-5-carboxylic acid tert-butyl ester,

cis-8-chloro-1-[4-(pyridazin-3-yloxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzo[e]azulene-5-carboxylic acid tert-butyl ester,

trans-8-chloro-5-methyl-1-[4-(pyridazin-3-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azulene,

cis-8-chloro-1-[4-(pyridin-2-yloxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzo[e]azulene-5-carboxylic acid tert-butyl ester,

cis-8-chloro-1-[4-(pyridin-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azulene,

cis-8-chloro-5-methyl-1-[4-(pyridin-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azulene,

trans-8-chloro-1-[4-(3-fluoro-pyridin-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azulene,

trans-8-chloro-1-[4-(3-fluoro-pyridin-2-yloxy)-cyclohexyl]-5-methyl-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azulene,

trans-8-Chloro-1-[4-(5-fluoro-pyridin-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azulene

trans-8-chloro-1-[4-(5-fluoro-pyridin-2-yloxy)-cyclohexyl]-5-methyl-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azulene,

trans-8-chloro-1-[4-(6-methyl-pyridin-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azulene,

trans-8-chloro-5-(2,2-difluoro-ethyl)-1-[4-(6-methyl-pyridin-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azulene,

trans-8-chloro-5-(2-fluoro-ethyl)-1-[4-(5-fluoro-pyridin-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azulene,

trans-8-chloro-5-ethyl-1-[4-(3-fluoro-pyridin-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azulene,

trans-8-chloro-5-ethyl-1-[4-(5-fluoro-pyridin-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azulene,

trans-8-chloro-5-ethyl-1-[4-(6-methyl-pyridin-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azulene, and

trans-8-Chloro-5-methyl-1-[4-(6-methyl-pyridin-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azulene.

One embodiment of the present invention relates to the use of a Via antagonist for the treatment and/or prevention of phase shift sleep disorders, wherein the Via antagonist is

Example 1.

One embodiment of the present invention is directed to a method for treating and/or preventing phase shift sleep disorders in a patient in need of such treatment, comprising administering to the patient a therapeutically effective amount of a Via antagonist of formula I as described herein.

One embodiment of the present invention relates to a method for treating and/or preventing phase shift sleep disorders in a patient in need of such treatment, said method comprising administering to said patient a therapeutically effective amount of a Via antagonist.

One embodiment of the present invention relates to a pharmaceutical composition comprising a Via antagonist in a pharmaceutically acceptable form.

One embodiment of the present invention relates to a pharmaceutical composition comprising a Via antagonist of formula I as described herein in a pharmaceutically acceptable form.

One embodiment of the present invention relates to a pharmaceutical composition comprising a Via antagonist of formula I as described herein in a pharmaceutically acceptable form for the treatment and/or prevention of phase shift sleep disorders.

One embodiment of the present invention relates to a pharmaceutical composition comprising a Via antagonist in a pharmaceutically acceptable form for the treatment and/or prevention of phase shift sleep disorders.

One embodiment of the present invention relates to a Via antagonist of formula I as described herein for use in the treatment and/or prevention of phase shift sleep disorders.

One embodiment of the present invention relates to a selective Via antagonist of formula I as described herein as Example 1 for use in the treatment and/or prevention of phase shift sleep disorders.

One embodiment of the invention relates to a selective Via antagonist of formula I as described herein as Example 1 for use in the treatment and/or prevention of jet lag.

One embodiment of the present invention relates to Via antagonists for use in the treatment and/or prevention of phase shift sleep disorders.

One embodiment of the present invention relates to Via antagonists for use in the preparation of a medicament for the treatment and/or prevention of phase shift sleep disorders.

One embodiment of the present invention relates to a Via antagonist of formula I as described herein for use in the preparation of a medicament for the treatment and/or prevention of phase shift sleep disorders.

One embodiment of the present invention relates to a selective Via antagonist of formula I as described herein for use in the treatment and/or prevention of phase shift sleep disorders.

One embodiment of the present invention relates to selective Via antagonists for use in the treatment and/or prevention of phase shift sleep disorders.

One embodiment of the present invention relates to selective Via antagonists for use in the preparation of medicaments for the treatment and/or prevention of phase shift sleep disorders.

One embodiment of the present invention relates to a selective Via antagonist of formula I as described herein for use in the preparation of a medicament for the treatment and/or prevention of phase shift sleep disorders.

One embodiment of the present invention relates to the use of a Via antagonist for the preparation of a medicament for the treatment and/or prevention of phase shift sleep disorders.

One embodiment of the present invention relates to the use of a Via antagonist of formula I as described herein for the preparation of a medicament for the treatment and/or prevention of phase shift sleep disorders.

Pharmaceutical composition

The compounds of formula I and their pharmaceutically acceptable salts can be used as medicines, for example in the form of pharmaceutical preparations. The pharmaceutical preparations can be administered orally, for example in the form of tablets, coated tablets, lozenges, hard and soft gelatin capsules, solutions, emulsions or suspensions. However, they can also be administered rectally, for example in the form of suppositories, or parenterally, for example in the form of injections.

The compounds of formula I and pharmaceutically acceptable salts thereof can be processed with pharmaceutically inert, inorganic or organic excipients for the preparation of tablets, coated tablets, dragees and hard gelatin capsules. Lactose, corn starch or derivatives thereof, talc, stearic acid or salts thereof, etc. can be used as such excipients for tablets, dragees and hard gelatin capsules. Excipients suitable for soft gelatin capsules are, for example, vegetable oils, waxes, fats, semisolid and liquid polyols, etc.

Excipients suitable for preparing solutions and syrups are, for example, water, polyols, sucrose, invert sugar, glucose, etc. Excipients suitable for injections are, for example, water, alcohols, polyols, glycerol, vegetable oils, etc. Excipients suitable for suppositories are, for example, natural or hardened oils, waxes, fats, semi-liquid or liquid polyols, etc.

In addition, the pharmaceutical preparations may contain preservatives, solubilizers, stabilizers, wetting agents, emulsifiers, sweeteners, colorants, flavorings, salts for varying the osmotic pressure, buffers, masking agents or antioxidants. They may also contain other therapeutically valuable substances.

Dosage can vary within a wide range and, of course, will be suitable for individual needs in each specific case. Usually, in the case of oral administration, a daily dosage of about 10 to 1000 mg/ people of the compound of Formula I should be suitable, but when necessary, the above-mentioned upper limit can also be exceeded.

Examples of compositions according to the present invention include, but are not limited to:

Example A

Tablets with the following composition are manufactured in the usual manner:

Table 1: Possible tablet compositions

Manufacturing process:

1. Mix ingredients 1, 2, 3 and 4 and granulate with purified water.

2. Dry the granules at 50°C.

3. Pass the granules through suitable grinding equipment.

4. Add ingredient 5 and mix for three minutes; compress on a suitable press.

Example B-1

Capsules with the following composition were prepared:

Table 2: Possible capsule composition

Manufacturing process:

1. Mix ingredients 1, 2 and 3 in a suitable mixer for 30 minutes.

2. Add ingredients 4 and 5 and mix for 3 minutes.

3. Fill into suitable capsules.

The compound of Formula I-III, lactose, and corn starch are first mixed in a mixer and then in a grinder. The mixture is returned to the mixer; talc is added and mixed thoroughly. The mixture is filled into suitable capsules, such as hard gelatin capsules, by machine.

Example B-2

Prepare soft gelatin capsules having the following composition:

Element mg/capsule Compounds of formula I 5 Yellow wax 8 hydrogenated soybean oil 8 Partially hydrogenated vegetable oil 34 soybean oil 110 total 165

Table 3: Possible soft gelatin capsule composition

Element mg/capsule gelatin 75 Glycerin 85% 32 Karion 83 8 (dry matter) Titanium dioxide 0.4 Iron oxide yellow 1.1 total 116.5

Table 4: Possible soft gelatin capsule compositions

Manufacturing Process

The compound of formula I is dissolved in the warm melt of the other ingredients and the mixture is filled into soft gelatin capsules of appropriate size. The filled soft gelatin capsules are processed according to the general procedures.

Example C

Prepare suppositories of the following composition:

Element mg/suppository Compounds of formula I 15 Suppository blocks 1285 total 1300

Table 5: Possible suppository compositions

Manufacturing Process

The suppository mass is melted in a glass or steel container, mixed thoroughly and cooled to 45°C. Subsequently, the finely powdered compound of formula I is added and stirred until it is completely dispersed. The mixture is poured into a suppository mold of appropriate size and left to cool; the suppositories are then removed from the mold and individually wrapped in wax paper or metal foil.

Example D

Prepare the following injection:

Element mg/injection Compounds of formula I 3 polyethylene glycol 400 150 Acetic acid Adjust to pH 5.0 Water for injection to 1.0ml

Table 6: Possible injection compositions

Manufacturing Process

The compound of formula I is dissolved in a mixture of polyethylene glycol 400 and water for injection (part). The pH is adjusted to 5.0 by acetic acid. The volume is adjusted to 1.0 ml by adding excess water. The solution is filtered, filled into a vial with an appropriate excess, and sterilized.

Example E

Create a sachet of the following composition:

Element mg/sachet Compounds of formula I 50 Lactose, finely powdered 1015 Microcrystalline cellulose (AVICEL PH 102) 1400 Sodium carboxymethyl cellulose 14 Polyvinylpyrrolidone K 30 10 magnesium stearate 10 Fragrance additives 1 total 2500

Table 7: Possible sachet compositions

Manufacturing Process

The compound of formula I is mixed with lactose, microcrystalline cellulose and sodium carboxymethylcellulose and granulated with a mixture of polyvinylpyrrolidone in water. The granules are mixed with magnesium stearate and flavoring additives and filled into sachets.

Example

1) Materials and methods

In all studies described below, male young adult C57BL/6J mice (4 weeks old at the start of the experiment) were used. Animals were exposed to a light-dark cycle (LD) of 12:12 (lights on from 8 h to 20 h geographic time; light intensity of 200 lux; dim red light of 5 lux during the dark period) in individual cages with ad libitum access to food and water and controlled temperature (22 ± 1 ° C) and humidity (55% ± 5%). The LD cycle is expressed as environmental clock time (zeitgeber times, ZT), where ZT0 represents lights on and ZT12 represents lights off. When animals were exposed to constant dark conditions (DD), circadian time (CT) was used as a time point reference, where CT12 corresponds to the onset of activity.

Characterization of Behavioral Circadian Responses to the Specific Via Antagonist, trans-8-chloro-5-methyl-1-(4-pyridin-2-yloxycyclohexyl)-4,6-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine (Example 1) in Mice (ZT12 and ZT0)

To assess whether there is a phase window of sensitivity to the Via receptor antagonist trans-8-chloro-5-methyl-1-(4-pyridin-2-yloxycyclohexyl)-4,6-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine (Example 1). A group of animals was treated at two time points (one during the day and one at night). Animals were exposed to LD cycling conditions (at least 10 days) to obtain a baseline of spontaneous activity rhythm. Four groups were selected: (Group 1) sham operation, in which the cannula was introduced into the oral cavity but no solution was delivered; (Group 2) no-load, and (Groups 2 and 3) administration of the Via receptor antagonist trans-8-chloro-5-methyl-1-(4-pyridin-2-yloxycyclohexyl)-4,6-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine (doses of 10 mg/kg and 30 mg/kg). Administration was performed at ZT/CT12 (lights off/activity on; sham, n=5; no-load n=5; 10 mg/kg n=5; and 30 mg/kg n=5) (oral administration). After the first manipulation at ZT/CT12, animals were immediately exposed to DD conditions and maintained for at least another 10 days for behavioral analysis, then re-adapted to a 12-12 LD cycle for a minimum of 10 days before being exposed to the second experimental manipulation at ZT/CT0 (lights on/activity offset; sham, n=3; vehicle, n=3; 10 mg/kg, n=5; and 30 mg/kg, n=5).

Characterization of Behavioral Circadian Responses to the Specific Via Antagonist, trans-8-chloro-5-methyl-1-(4-pyridin-2-yloxycyclohexyl)-4,6-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine (Example 1) in Mice (ZT18 and ZT6)

A new group of animals was treated at two additional time points different from the previous experiment; ZT/CT6 (6 h after lights on/mid-rest period; sham, n=3; vehicle n=4; and 30 mg/kg n=5) and ZT/CT18 (6 h after lights off/mid-active period; sham, n=3; vehicle n=4; and 30 mg/kg n=5). For this experiment (time points ZT/CT6 and ZT/CT18), trans-8-chloro-5-methyl-1-(4-pyridin-2-yloxycyclohexyl)-4,6-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine was administered only at a dose of 30 mg/kg. Therefore, three groups were selected: (Group 1) sham operation, (Group 2) vehicle administration, and (Group 3) trans-8-chloro-5-methyl-1-(4-pyridin-2-yloxycyclohexyl)-4,6-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine (30 mg/kg). Immediately after the operation, the animals were exposed to DD conditions and maintained for at least another 10 days for behavioral analysis before being acclimated to a 12-12 LD cycle for a minimum of 10 days.

Effects of the V1a antagonist trans-8-chloro-5-methyl-1-(4-pyridin-2-yloxycyclohexyl)-4,6-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine on photosynchronization of the clock in mice

To assess the effects of the Via antagonist trans-8-chloro-5-methyl-1-(4-pyridin-2-yloxycyclohexyl)-4,6-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine on light synchronization of behavioral circadian rhythms, a 6-h phase-shifted jet lag protocol was used. After being placed on a 12-h/12-h light/dark cycle for at least 10 days, mice were exposed to a 6-h phase shift of the light/dark cycle (the first "jet lag" test, premature shift) and were exposed to either no vehicle or 30 mg/kg of the Via antagonist trans-8-chloro-5-methyl-1-(4-pyridin-2-yloxycyclohexyl)-4,6-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine. On the day of shifting, the lights-off time was phase shifted by 6 h. Three groups of animals were used: a sham operation group (Group 1, n = 3), animals in which a cannula was introduced into the oral cavity but no solution was delivered; a vehicle group (Group 2, n = 4), which received no vehicle, and a 30 mg/kg trans-8-chloro-5-methyl-1-(4-pyridin-2-yloxycyclohexyl)-4,6-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine group (Group 3, n = 5). All operations were performed at ZT6 (6 hours after lights on) of the last LD cycle before a 6-hour phase advance.

In a second experiment, naive animals were used to evaluate the second jet lag test. Therefore, a new group of male mice was used to test the effect of 30 mg/kg trans-8-chloro-5-methyl-1-(4-pyridin-2-yloxycyclohexyl)-4,6-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine on resynchronization of the 6-h phase advance of the LD cycle. At ZT/CT6, animals that had been on a 12 h/12 h LD cycle (for at least 10 days) were exposed to a 6-h phase advance of the LD cycle and to either vehicle or 30 mg/kg of the Via antagonist trans-8-chloro-5-methyl-1-(4-pyridin-2-yloxycyclohexyl)-4,6-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine. Similar to the previous experiments, three groups of animals were used: a sham operation group (Group 1, n=3), animals in which a cannula was introduced into the mouth but no solution was delivered; a vehicle group (Group 2, n=4), which received vehicle, and a 30 mg/kg trans-8-chloro-5-methyl-1-(4-pyridin-2-yloxycyclohexyl)-4,6-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine group (Group 3, n=5).

Behavior recording and analysis

Spontaneous activity rhythms were measured using infrared detectors on the cage top and/or on a running wheel. Activity data were displayed as actograms using the program ClockLab (Actimetrics, Evanston, IL, USA) and analyzed for the following parameters:

Phase shift: Phase shift was measured by fitting two regression lines to the onset of spontaneous activity 10 days before and 10 days after the procedure (ClockLab; Actimetrics, Evanston, IL, USA). A computer algorithm was used to determine the onset of the main period of daily spontaneous activity (ClockLab). Data collected one day after the procedure (sham, empty vehicle, or drug administration) were excluded from the line fit to ensure that the circadian phase was in steady state. Phase shift was defined as the difference (in hours) between the pre-stimulus line and the post-stimulus line. The magnitude of the phase shift was fitted by linear regression to the onset, taking into account 10 cycles before compound administration (LD) and 10 cycles after compound administration (DD; Aschoff Type II procedure to evaluate phase shifts; Mrosovsky, 1996b) when the animals were released in constant darkness.

Period: The intrinsic period was determined by analyzing the χ2 periodogram (ClockLab).

Alpha: The duration of the active period (alpha) before and after treatment was also calculated. For this, the program ClockLab was used to determine the onset and offset of the activity (in both LD and DD). The difference (in hours) between these two reference points represents alpha, or the duration of the active period. The onset and offset of the activity were automatically identified using the ClockLab software.

Activity profile: Activity data were collected in 30-minute bins and plotted as waveforms using ClockLab, averaging the 6-h activity of the animals after the procedure (drug administration day). The group average waveform of activity was obtained from the 6-h activity of each animal after the procedure.

Readaptation (jet lag test): For a 6 h phase shift (jet lag protocol), the number of days required for complete readaptation after the shift was quantified, taking into account both the onset and the offset of activity. The onset and offset of activity were automatically identified using ClockLab software. Therefore, when the onset of activity occurred at a new time of ±30 minutes after the lights went out, it was considered to be completely resynchronized with the new LD cycle after a 6-hour phase shift of the time of lights out. For each mouse, the number of days required for resynchronization was calculated (short-term cycles). The speed of readaptation was assessed daily, and the onset of activity in hours and its advance were calculated. In order to evaluate the effect of the manipulation on locomotion, the changes in spontaneous activity from 0 to 6 h after each treatment were quantified at intervals of 30 min (see activity curve determination).

Statistical analysis

All values in the experiments are mean ± SEM. Statistical comparisons were performed using one-way analysis of variance (ANOVA) for independent (phase shift, period, readaptation) and repeated measures (activity curve, alpha) and followed by post hoc Fisher's (LSD) test. For all experiments, the critical value of significance was p < 0.05. The statistical software package Statistica (version 8.0) was used.

result

Phase shift, period, alpha, and activity curves of behavioral circadian rhythms in response to trans-8-chloro-5-methyl-1-(4-pyridin-2-yloxycyclohexyl)-4,6-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine (Example 1) administered at ZT/CT12 and ZT/CT0

For the first part of the experiment at the two time points tested (ZT/CT12 and ZT/CT0), two doses of the Via receptor antagonist trans-8-chloro-5-methyl-1-(4-pyridin-2-yloxycyclohexyl)-4,6-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine were used, 10 mg/kg and 30 mg/kg.

Assessment of behavioral phase shifts following single administration of Via antagonist at 10 or 30 mg/kg revealed no effect on circadian phase (ZT/CT12, _F3,16 = 0.55, p = 0.65; ZT/CT0, _F3,12 = 0.14, p = 0.93; Figure 1).

No significant changes in locomotor activity were observed after administration of the Via antagonist at 10 mg/kg or 30 mg/kg. Intrinsic locomotor activity was also unchanged in animals treated at ZT/CT12 and ZT/CT0 (ZT/CT12, F _3,16 = 0.4, p = 0.72; ZT/CT0, F _3,12 = 0.3, p = 0.8; Figure 2).

Although animals treated at ZT/CT12 showed a change in alpha after treatment (time before vs. time after, F _1,16 = 34.5, p = 0.00002), no differences between groups were observed (each group, F _3,16 = 0.08, p = 0.96). Mice that received the Via antagonist at ZT/CT0 at both 10 and 30 mg/kg doses also showed an increase in alpha (time before vs. time after, F _1,12 = 81.8, p = 0.000001). Although alpha increased for almost 2 hours in Via antagonist-treated animals, the effect was not statistically significant (F _3,12 = 1.48, p = 0.26).

Phase shift, period, alpha, and activity curves of behavioral circadian rhythms in response to trans-8-chloro-5-methyl-1-(4-pyridin-2-yloxycyclohexyl)-4,6-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine (Example 1) administered at ZT/CT18 and ZT/CT6

Because no effects on phase shift, period, and alpha were found at the 10 mg/kg dose, for the second part of the experiment, animals received either vehicle or trans-8-chloro-5-methyl-1-(4-pyridin-2-yloxycyclohexyl)-4,6-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine at ZT/CT18 (mid-active period) or ZT/CT6 (mid-rest period).

At ZT/CT18, animals were treated with either sham, vehicle, or a 30 mg/kg dose of trans-8-chloro-5-methyl-1-(4-pyridin-2-yloxycyclohexyl)-4,6-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine. Assessment of behavioral phase shifts following a single 30 mg/kg dose of the Via antagonist showed no significant effect on circadian phase at ZT/CT18 (F _2,9 =0.13, p=0.87; nor at ZT/CT6 (F _2,9 =0.96, p=0.41; Figure 1).

At ZT/CT18 and ZT/CT6, the intrinsic period of movement was also not significantly different between the groups (ZT/CT18, _F2,9 = 0.1, p = 0.9; ZT/CT6, _F2,9 = 3, p = 0.12; Figure 2).

Animals treated at ZT/CT18 or ZT/CT6 showed changes in alpha after treatment (ZT/CT18, time before vs. time after, _F1,9 = 26.13, p = 0.0006; ZT/CT6 before vs. after, _F1,9 = 52.31 p = 0.00004), however, no differences were observed between the groups (ZT/CT18 groups, _F2,9 = 1.35, p = 0.3; ZT/CT6, _F2,9 = 2.4, p = 0.14).

Behavioral Readaptation to the 6-h Phase Shift of the LD Cycle: Effect of Administration of trans-8-chloro-5-methyl-1-(4-pyridin-2-yloxycyclohexyl)-4,6-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine (Example 1) (Jet Lag Test 1)

It was demonstrated that a specific V1a antagonist affects light synchronization in animals exposed to an experimental phase-shifting protocol.

Therefore, on the last day of the first LD cycle, animals exposed to a 6-h phase shift of the light-dark cycle at ZT6 (6 h after lights on) (sham, vehicle, and Via antagonist groups) were treated with either a vehicle control or 30 mg/kg of a Via antagonist. Animals from the sham-controlled group took 5.66 ± 0.27 days to adapt to the new LD cycle. Animals receiving a vehicle dose took 5.75 ± 0.54 days to readapt to the new LD cycle. In contrast, mice treated with 30 mg/kg of the Via receptor antagonist trans-8-chloro-5-methyl-1-(4-pyridin-2-yloxycyclohexyl)-4,6-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine showed complete behavioral readaptation after the 6-h phase shift in just 3.2 ± 0.33 days. This effect was statistically significant (F _2,9 = 10.1, p = 0.005; Figure 3). In the group receiving the Via antagonist trans-8-chloro-5-methyl-1-(4-pyridin-2-yloxycyclohexyl)-4,6-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine, the onset of activity showed a much faster daily regulation of the LD cycle during the first 5 days after the shift (F _2,9 = 25, p = 0.0002). The spontaneous activity induced by the manipulation did not differ significantly between the groups (F _2,9 = 1.25, p = 0.32).

The first experiment was repeated under the same experimental conditions, but using new naive mice (young adult male mice not previously exposed to any type of manipulation).

Naive animals from the sham control group took 5.33 ± 0.88 days to adapt to the new LD cycle. Animals that received vehicle required 5.5 ± 0.64 days to readapt to the new LD cycle. Similar to the first phase-shift experiment, mice treated with 30 mg/kg of the Via antagonist trans-8-chloro-5-methyl-1-(4-pyridin-2-yloxycyclohexyl)-4,6-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine showed complete behavioral readaptation in only 3.4 ± 0.24 days after the 6-h phase shift. This effect was statistically significant (F = _4.99 , p = 0.03; Figure 4). In the group treated with the Via antagonist trans-8-chloro-5-methyl-1-(4-pyridin-2-yloxycyclohexyl)-4,6-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine, the onset of activity showed a much faster daily regulation of the LD cycle during the first 4 days after the shift ( _F2,9 = 14, p = 0.001).

The three experimental manipulations (sham, vehicle, and compound administration) induced increases in locomotion that were not significantly different between the groups ( _F2,9 = 2.91, p = 0.10).

Both experiments indicate that the Via antagonist trans-8-chloro-5-methyl-1-(4-pyridin-2-yloxycyclohexyl)-4,6-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine accelerates the readaptation of the circadian clock to the new LD cycle.

2) Materials and Methods

Preparation of test compounds

Test compounds were first dissolved in DMSO at a concentration of 10 mM and subsequently diluted in assay buffer to a maximum final concentration of 0.2% DMSO.

Radioligand binding assay membrane preparation

Recombinant vasopressin V _1a (human: hV _1a ; mouse: mV _1a ) and V _1b (human: hV _1b ) were independently expressed in human embryonic kidney (HEK) cells using transient transfection. Cell membranes expressing various receptors were prepared by homogenizing the cell culture pellet (using a Polytron homogenizer) in lysis buffer (50 mM HEPES, 1 mM EDTA, 10 mM MgCl ₂ , pH 7.4, with protease inhibitors) and centrifuging the resulting suspension at 500 g for 20 min at 4°C. The supernatant was then centrifuged again at 43,000 g for 1 hour at 4°C. The resulting pellet was resuspended in lysis buffer and 10% sucrose. Protein concentration was determined, and membranes were aliquoted and stored at -80°C until further use.

Radioligand binding assay to evaluate the affinity of test compounds for vasopressin receptors from different species

All radioligand binding assays were performed in 96-well plates in the presence of 1 nM ^3H -AVP (vasopressin) radioligand and 10 concentrations of test compound (27 μM to 1 nM or 2.7 μM to 0.1 nM for hV _1a binding). Compound dilutions were performed in assay buffer using a Beckman Biomek 2000 laboratory automation workstation. Nonspecific binding was limited using 4.2 μM cold vasopressin. Each well contained membrane protein (at varying concentrations) and 0.5 mg of Ysi-poly-1-lysine SPA beads (used only for human receptor SPA assays, not for mouse V 1a filtration assays) in a final volume of 200 μl of buffer A containing: Tris 50 mM, NaCl 120 mM, KCl 5 mM, CaCl 2 2 mM, MgCl ₂ 10 mM, 1 mM _Cysteine (pH 7.4). All assays were carried out in duplicate and repeated at least twice. The assay plate was incubated at room temperature for 1 hour, then centrifuged (for people's receptor (SPA)) or filtered (for mouse receptor). The filtration assay was terminated by rapid filtration through a GF/C filter under vacuum, pre-soaked for 5 min with the buffer B containing Tris 50 mM, MgCl 5 mM, 0.1% BSA (pH 7.4), and then washed with 5 x 0.4 ml of ice-cold buffer B. For both SPA and filter plates, a Packard Topcount scintillation counter was used to measure the combined part. The single homogeneous colony of the saturation binding experiment indication binding site for each assay was labeled.

Calculation of radioligand binding data.

The CPM values of two copies of a certain concentration of competing compound were averaged (y1) and % specific binding was calculated, (((y1-nonspecific)/(total binding-nonspecific)) x 100). XLfit (a curve fitting program that uses the Levenburg Marquardt algorithm to iteratively plot the data) was used to plot the % specific binding. The single site competition assay equation used was y = A + ((B-A)/(1+((x/C) ^D ))), where y is % specific binding, A is the minimum y, B is the maximum y, C is IC50, x is the log ₁₀ of the concentration of the competing compound, and D is the slope of the curve (Hill Coefficient). From these curves, IC50 (the inhibitory concentration at which 50% of the specific binding of the radioligand is displaced) and the Hill coefficient were determined. The affinity constant (Ki) was calculated using the Cheng-Prussoff equation: Ki = (IC50/1 + ([L]/Kd), where [L] is the concentration of the radiolabel and Kd is the equilibrium dissociation constant of the radiolabel. Ki is also expressed logarithmically as pKi.

result

Table 1. Affinity of Example 1 at human and mouse _V1a and human _V1b receptors in radioligand binding assays

Ki[nM] people 0.7±0.1(5) >27,000(2) mice 77±2(3) n.t.

Data are expressed as pKi±SEM(n) or Ki[nM]±SEM(n), where pKi is -Log ₁₀ of affinity constant (Ki)[M], SEM is standard error of the mean, and n is the number of assays (nt is not tested).

Example 1:

trans-8-chloro-5-methyl-1-(4-pyridin-2-yloxycyclohexyl)-4,6-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine (trans-8-chloro-5-methyl-1-[4-(pyridin-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoazulene)

Example 1.

Compounds of formula I, in particular Example 1, can be prepared as described in PCT application WO 2010/060836.

Attached photos

Figure 1: (Top) Representative activity graphs of spontaneous activity in different groups treated at ZT/CT12 (activity onset) or ZT0/CT0 (activity offset). The dots in the activity graphs indicate the time of manipulation, while the yellow and black bars in the upper part indicate the light and dark periods, respectively. (Middle) Mean ± SEM of the rhythmic phase shift of spontaneous activity in mice treated at ZT/CT12 (activity onset; sham, n=5; vehicle n=5; 10 mg/kg n=5; 30 mg/kg n=5) or ZT/CT0 (activity offset; sham, n=3; vehicle n=3; 10 mg/kg n=5; 30 mg/kg n=5). (Bottom) Mean (±SEM) of rhythmic phase shifts in spontaneous activity of mice treated (under different experimental conditions) at ZT/CT18 (sham, n=3; vehicle n=4; 30 mg/kg n=5) or ZT/CT6 (sham, n=3; vehicle n=4; 30 mg/kg n=5).

Figure 2: Mean (±SEM) of the intrinsic period of spontaneous activity of mice under constant dark conditions after each experimental treatment at ZT/CT12 (activity onset; sham, n=5; vehicle n=5; 10 mg/kg n=5; 30 mg/kg n=5), ZT0/CT0 (activity offset; sham, n=3; vehicle n=3; 10 mg/kg n=5; 30 mg/kg n=5), ZT/CT18 (midnight; sham, n=3; vehicle n=4; 30 mg/kg n=5) and ZT/CT6 (noon; sham, n=3; vehicle n=4; 30 mg/kg n=5).

Figure 3: (Top) Representative activity graphs of spontaneous activity of different groups treated at ZT/CT6 (intermediate rest period) and exposed to a 6 h phase advance of the LD cycle. The dots in the activity graphs indicate the time of manipulation, while the yellow and black bars in the upper and lower parts indicate the light and dark cycles, respectively. (Middle) The rate (number of days) of fully synchronized readaptation to the new LD cycle is shown (sham n=3, vehicle n=4, 30 mg/kg n=5). The adaptation speed was assessed daily. From day 1 to day 5, the group treated with 30 mg/kg of the Via antagonist trans-8-chloro-5-methyl-1-(4-pyridin-2-yloxycyclohexyl)-4,6-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine showed significantly faster readaptation.

Figure 4: (Top) Representative activity graphs of spontaneous activity of different groups of naive mice treated at ZT/CT6 (intermediate rest period) and exposed to a 6 h phase advance of the LD cycle. The dots in the activity graphs indicate the time of manipulation, while the yellow and black bars in the upper and lower parts indicate the light and dark cycles, respectively. (Middle) The rate (number of days) of fully synchronized readaptation to the new LD cycle is shown (sham n=3, vehicle n=4, 30 mg/kg n=5). The adaptation speed was assessed daily. From day 1 to day 4, the group treated with 30 mg/kg of the Via antagonist trans-8-chloro-5-methyl-1-(4-pyridin-2-yloxycyclohexyl)-4,6-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine showed significantly faster readaptation.

Claims (9)

1. Use of a V1a antagonist in the preparation of a medicament for the treatment and/or prevention of phase shift sleep disorders, wherein said V1a antagonist is a compound of formula I. in ^R1 is phenyl, naphthyl, pyridyl, pyrimidinyl, pyridazinyl, or pyrazinyl, which is unsubstituted or substituted by one or more substituents independently selected from A. ^R2 is H, _C1-12 -alkyl, which is unsubstituted or substituted with one or more OH groups or halogens. -( _CH₂ ) _q - ^Ra , where ^Ra is a pyridyl group. -C(O)-C _1-12 -alkyl, -C(O)(CH ₂ ) _q NR ⁱ R ⁱⁱ ， -C(O)OC _1-12 -alkyl, -S(O) ₂ -C _1-12 -alkyl, or -S(O) ₂ NR ⁱ R ⁱⁱ ， ^Ri and ^Rii are each independently H or _C1-12 -alkyl. q is 1, A is a halogen, cyano, _C1-7 -alkyl, halo- _C1-7 -alkyl, _C1-7 -alkoxy, or halo- _C1-7 -alkoxy group. ^R3 is Cl or F. Or its medicinal salt. 2. The use according to claim 1, wherein the V1a antagonist is a compound of formula I, wherein ^R1 is phenyl, naphthyl, pyridyl, pyrimidinyl, pyridazinyl, or pyrazinyl, each of which is unsubstituted or substituted by one or more substituents independently selected from A; and A is a halogen, cyano, _C1-7 -alkyl, halogenated- _C1-7 -alkyl, or _C1-7 -alkoxy group. 3. The use according to any one of claims 1-2, wherein the V1a antagonist is a compound of formula I, wherein ^R1 is a phenyl group. 4. The use according to any one of claims 1-3, wherein the V1a antagonist is a compound of formula I, wherein ^R1 is pyridyl. 5. The use according to any one of claims 1-4, wherein the V1a antagonist is a compound of formula I, wherein ^R2 is ■H, ■C _1-12 -alkyl, which is unsubstituted or substituted with one or more OH groups. ■-( _CH₂ ) _q - ^Ra , where ^Ra is a pyridyl group and q is 1. ■-C(O)-C _1-12 -alkyl, ■-C(O)( _CH₂ ) _qNRiRi , where ^Ri and ^Ri are each independently C₁ _-12 ^{- alkyl} , and q is 1. ■-C(O)OC _1-12 -alkyl, ■-S(O) ₂ -C _1-12 -alkyl, or ■-S(O) ₂ NR ⁱ R ⁱⁱ , wherein ^Ri and ^Ri are each independently _C1-12 -alkyl. 6. The use according to any one of claims 1-5, wherein the V1a antagonist is a compound of formula I, wherein ^R2 is methyl. 7. The use according to any one of claims 1-6, wherein the V1a antagonist is a compound of formula I, wherein ^R3 is Cl. 8. The use according to any one of claims 1-7, wherein the V1a antagonist is a compound of formula I, wherein the compound is selected from the group consisting of: trans-8-chloro-1-(4-phenoxy-cyclohexyl)-4H,6H-2,3,5,10b-tetraaza-benzoxazine-tert-butyl 5-carboxylate trans-8-chloro-1-(4-phenoxy-cyclohexyl)-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoxazine hydrochloride, trans-8-chloro-5-methyl-1-(4-phenoxy-cyclohexyl)-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azine, trans-1-[8-chloro-1-(4-phenoxy-cyclohexyl)-4H,6H-2,3,5,10b-tetraaza-benzoxazine-5-yl]-ethyl ketone, trans-8-chloro-5-methanesulfonyl-1-(4-phenoxy-cyclohexyl)-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoxazine, trans-2-[8-chloro-1-(4-phenoxy-cyclohexyl)-4H,6H-2,3,5,10b-tetraaza-benzoxazine-5-yl]-ethanol, trans-8-chloro-5-isopropyl-1-(4-phenoxy-cyclohexyl)-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoxazine, trans-8-chloro-1-(4-phenoxy-cyclohexyl)-4H,6H-2,3,5,10b-tetraaza-benzoxazine-5-sulfonic acid dicarboxamide, trans-8-chloro-1-(4-phenoxy-cyclohexyl)-5-pyridin-2-ylmethyl-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoxazine, trans-1-[8-chloro-1-(4-phenoxy-cyclohexyl)-4H,6H-2,3,5,10b-tetraaza-benzoxazine-5-yl]-2-dimethylamino-ethyl ketone, trans-8-fluoro-1-(4-phenoxy-cyclohexyl)-4H,6H-2,3,5,10b-tetraaza-benzo[e]azine-tert-butyl 5-carboxylate trans-8-fluoro-5-methyl-1-(4-phenoxy-cyclohexyl)-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoxazine, cis-8-chloro-1-(4-phenoxy-cyclohexyl)-4H,6H-2,3,5,10b-tetraaza-benzoxazine-tert-butyl 5-carboxylate cis-8-chloro-5-methyl-1-(4-phenoxy-cyclohexyl)-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoxazine, trans-8-chloro-1-[4-(4-fluoro-phenoxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzoxazine-5-carboxylic acid tert-butyl ester, trans-8-chloro-1-[4-(4-fluoro-phenoxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azine hydrochloride, trans-8-chloro-1-[4-(4-fluoro-phenoxy)-cyclohexyl]-5-methyl-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoxazine, trans-8-chloro-1-[4-(4-cyano-phenoxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzo[e]azine-tert-butyl 5-carboxylate, trans-4-[4-(8-chloro-5-methyl-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azine-1-yl)-cyclohexyloxy]benzylnitrile, trans-8-chloro-1-[4-(4-trifluoromethyl-phenoxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzo[e]azine-tert-butyl carboxylate, trans-8-chloro-1-[4-(3-chloro-phenoxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzoxazine-5-carboxylic acid tert-butyl ester, trans-8-chloro-1-[4-(3-chloro-phenoxy)-cyclohexyl]-5-methyl-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoxazine, trans-8-chloro-1-[4-(3-methoxy-phenoxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzoxazine-5-carboxylic acid tert-butyl ester, trans-8-chloro-1-[4-(3-methoxy-phenoxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azine hydrochloride, trans-8-chloro-1-[4-(3-methoxy-phenoxy)-cyclohexyl]-5-methyl-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoxazine, trans-3-[4-(8-chloro-5-methyl-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoxazine-1-yl)-cyclohexyloxy]-benzylnitrile, trans-8-chloro-1-(4-m-tolyloxy-cyclohexyl)-4H,6H-2,3,5,10b-tetraaza-benzo[e]azine-tert-butyl 5-carboxylate trans-8-chloro-1-(4-m-tolyloxy-cyclohexyl)-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azine hydrochloride, trans-8-chloro-5-methyl-1-(4-m-tolyloxy-cyclohexyl)-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azine, trans-1-[4-(3-tert-butyl-phenoxy)cyclohexyl]-8-chloro-4H,6H-2,3,5,10b-tetraaza-benzo[e]azine-5-carboxylic acid tert-butyl ester, trans-8-chloro-1-[4-(2-fluoro-phenoxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzo[e]azine-5-carboxylic acid tert-butyl ester, trans-8-chloro-1-[4-(2-fluoro-phenoxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azine hydrochloride, trans-8-chloro-1-[4-(2-fluoro-phenoxy)-cyclohexyl]-5-methyl-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azine, trans-8-chloro-1-[4-(2-cyano-phenoxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzoxazine-tert-butyl 5-carboxylate, trans-2-[4-(8-chloro-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azine-1-yl)-cyclohexyloxy]-benzylnitrile hydrochloride, trans-2-[4-(8-chloro-5-methyl-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoxazine-1-yl)-cyclohexyloxy]-benzylnitrile, trans-8-chloro-1-(4-o-tolyloxycyclohexyl)-4H,6H-2,3,5,10b-tetraaza-benzo[a]azine-tert-butyl 5-carboxylate trans-8-chloro-5-methyl-1-(4-o-tolyloxy-cyclohexyl)-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoxazine, trans-8-chloro-1-[4-(3,5-difluoro-phenoxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzoxazine-5-carboxylic acid tert-butyl ester, trans-8-chloro-1-[4-(3,5-difluoro-phenoxy)-cyclohexyl]-5-methyl-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoxazine, trans-8-chloro-1-[4-(naphthyl-2-yloxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzoxazine-5-carboxylic acid tert-butyl ester, trans-8-chloro-5-methyl-1-[4-(naphth-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoxazine, trans-8-chloro-1-[4-(pyridin-3-yloxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzoxazine-5-carboxylic acid tert-butyl ester, trans-8-chloro-5-methyl-1-[4-(pyridin-3-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoxazine, trans-8-chloro-1-[4-(5-chloro-pyridin-3-yloxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzoxazine-5-carboxylic acid tert-butyl ester, trans-8-chloro-1-[4-(5-chloro-pyridin-3-yloxy)-cyclohexyl]-5-methyl-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoxazine, trans-8-chloro-1-[4-(pyridin-2-yloxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzoxazine-5-carboxylic acid tert-butyl ester, trans-8-chloro-1-[4-(pyridin-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoxazine hydrochloride, trans-8-chloro-5-methyl-1-[4-(pyridin-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzoxazine, trans-8-chloro-1-[4-(6-chloro-pyridin-2-yloxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzo[e]azine-5-carboxylic acid tert-butyl ester, trans-8-chloro-1-[4-(6-chloro-pyridin-2-yloxy)-cyclohexyl]-5-methyl-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azine, trans-8-chloro-1-[4-(5-chloro-pyridin-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azine hydrochloride, trans-8-chloro-1-[4-(5-chloro-pyridin-2-yloxy)-cyclohexyl]-5-methyl-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azine, trans-8-chloro-1-[4-(pyrimidin-2-yloxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzo[e]azine-5-carboxylic acid tert-butyl ester, cis-8-chloro-1-[4-(pyrimidin-2-yloxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzo[e]azine-5-carboxylic acid tert-butyl ester, trans-8-chloro-5-methyl-1-[4-(pyrimidin-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azine, trans-8-chloro-1-[4-(pyrazin-2-yloxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzo[e]azine-5-carboxylic acid tert-butyl ester, trans-8-chloro-5-methyl-1-[4-(pyrazin-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azine, trans-8-chloro-1-[4-(pyrimidin-4-yloxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzo[e]azine-5-carboxylic acid tert-butyl ester, trans-8-chloro-5-methyl-1-[4-(pyrimidin-4-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azine, trans-8-chloro-1-[4-(pyridazin-3-yloxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzo[e]azine-5-carboxylic acid tert-butyl ester, cis-8-chloro-1-[4-(pyridazin-3-yloxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzo[e]azine-5-carboxylic acid tert-butyl ester, trans-8-chloro-5-methyl-1-[4-(pyridazin-3-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azine, cis-8-chloro-1-[4-(pyridin-2-yloxy)-cyclohexyl]-4H,6H-2,3,5,10b-tetraaza-benzo[e]azine-5-carboxylic acid tert-butyl ester, cis-8-chloro-1-[4-(pyridin-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azine, cis-8-chloro-5-methyl-1-[4-(pyridin-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azine, trans-8-chloro-1-[4-(3-fluoro-pyridin-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azine, trans-8-chloro-1-[4-(3-fluoro-pyridin-2-yloxy)-cyclohexyl]-5-methyl-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azine, trans-8-chloro-1-[4-(5-fluoro-pyridin-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azine trans-8-chloro-1-[4-(5-fluoro-pyridin-2-yloxy)-cyclohexyl]-5-methyl-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azine, trans-8-chloro-1-[4-(6-methylpyridin-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azine, trans-8-chloro-5-(2,2-difluoro-ethyl)-1-[4-(6-methyl-pyridin-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azine, trans-8-chloro-5-(2-fluoro-ethyl)-1-[4-(5-fluoro-pyridin-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azine, trans-8-chloro-5-ethyl-1-[4-(3-fluoro-pyridin-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azine, trans-8-chloro-5-ethyl-1-[4-(5-fluoro-pyridin-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azine, trans-8-chloro-5-ethyl-1-[4-(6-methylpyridin-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azine, and trans-8-chloro-5-methyl-1-[4-(6-methylpyridin-2-yloxy)-cyclohexyl]-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azine. 9. The use according to any one of claims 1-8, wherein the V1a antagonist is