CN1496354A

CN1496354A - Quinoline, isoquinoline and phthalazine derivatives as antagonists of gonadotropin-releasing hormone

Info

Publication number: CN1496354A
Application number: CNA028063597A
Authority: CN
Inventors: P; P·施特雷尔克; P·德勒舍尔; U·比曼; N·史梅斯; P·穆恩; -; H·赫斯-施通普; R·屈内; E·京特; ��Ī��; E·波利莫罗普洛斯; A·M·特拉克
Original assignee: Zentaris AG
Current assignee: Aeterna Zentaris GmbH
Priority date: 2001-02-21
Filing date: 2002-02-21
Publication date: 2004-05-12
Also published as: RU2003128647A; CA2438709A1; NO20033698L; JP2004528298A; MXPA03007348A; BR0207451A; WO2002066437A1; IL157303A0; AU2002247737B2; NO325675B1; SK11622003A3; PL367285A1; NZ527597A; RU2280031C2; CZ20032514A3; BG108165A; HUP0303219A2; EP1362034A1; NO20033698D0

Abstract

The invention relates to new quinoline, isoquinoline and phthalazine derivatives as antagonists of the gonadotropin-releasing hormone.

Description

Quinoline, isoquinoline 99.9 and phthalazine derivatives as antagonists of gonadotropin-releasing hormone

Gonadotropin releasing hormone (GnRH) is a kind of like this hormone, and it mainly but be not synthetic by the hypothalamus neurocyte in Mammals exclusively is transported to hypophysis via portal vein, is released in the gonadotroph in the mode of being regulated.By the interaction that---has 7 membrane spaning domains---with its acceptor, GnRH is by second messenger's inositol-1,4,5-triphosphoric acid and Ca ²⁺Ion stimulates the generation and the release of gonad-stimulating hormone.Stimulate the amphoteric sex steroid to produce and the sexual cell maturation by GnRH excretory gonad-stimulating hormone lutropin (LH) and follicle stimulating hormone (FSH).Except GnRH (being also referred to as GnRH1), also has the GnRH of two kinds of forms, i.e. GnRH2 and 3.

The GnRH acceptor is used as the pharmacology target some depending in the disease that the functionality hormone produces, for example mammary cancer, endometriosis and fibroma uteri before prostate cancer, the menopause.In these diseases, can successfully adopt super-agonists or the super-agonist of GnRH.Definite, male contraception is united the male hormone that substitutes dosage, constitutes further possible indication.

Compare with super-agonists, an advantage of GnRH antagonist is their instant effects in the retardance gonadotrophin secretion.Super-agonists causes the overstimulation of hypophysis at first, and the latter causes that the secretion of gonad-stimulating hormone and sex steroid increases.

This hormone response only have because of the desensitization of GnRH acceptor density and decrement regulating effect certain postpone after just termination.The GnRH super-agonists---is united---therefore by oneself and with testosterone can not suppress the generation of mankind spermatozoon effectively, thereby is not suitable for male contraception.In contrast, peptide class GnRH antagonist---definite male hormone with alternative dosage is united---can cause the significant oligospermia of the male sex.

But, peptide class GnRH antagonist has a lot of shortcomings.For example their activity is significantly less than super-agonists, so must give higher dosage.Their oral administration biaavailability is also low, so that they need drug administration by injection.Injection causes that then tolerance reduces repeatedly.And, compare not only required great effort gas but also take fund of peptide class GnRH antagonist synthetic with non-peptide compound.

Quinoline as non-peptide class GnRH antagonist for example is disclosed among the WO 97/14682.But so far still nothing but peptide class GnRH antagonist put on market.

The object of the present invention is to provide novel GnRH antagonist, they are better than known peptides, have represented effective alternative of known non-peptide compound.Novel GnRH antagonist should have very high activity and very high oral administration biaavailability.In addition, they should be synthesized simply, and cost is low as much as possible.

This purpose is realized by general formula (1) compound:

Wherein

R1 (a) is acyl group-CO-R11 or CN; wherein R11 is that saturated, unsaturated, ring-type is or/and the organic atomic group of (mixing) aromatics is definite in having the straight or branched alkyl chain of 1-10 C atom, perhaps phenyl, furans or thienyl; replaced by alkyl or halogen atom alternatively and

(b) carboxylic acid ester groups-CO-OR12 or formamido--CO-NR12NR13 or group-SO _x-R12---wherein x=0,1 or 2---or-SO ₂-NR12NR13, wherein R12 is that saturated, unsaturated, ring-type is or/and the organic atomic group of (mixing) aromatics, be that definitely the straight or branched alkyl chain with 1-10 C atom, the aralkyl with 7-20 C atom---wherein the aryl atomic group can be replaced or the phenyl atomic group by alkyl or halogen atom alternatively---are replaced by alkyl or halogen atom alternatively, R13 can be hydrogen atom or the straight or branched alkyl chain with 1-10 C atom, perhaps

(c) be group-A-NR14-CO-NR15R16, wherein A is the alkylidene group with 1-4 C atom, definitely is 1 C atom, alternatively by C ₁-C ₆-alkyl, carbonyl, Sauerstoffatom or group-SO _x-replace, x=0 wherein, 1 or 2, R14 and R15 are hydrogen atom or the straight or branched alkyl chain with 1-10 C atom independently of one another, R16 is the straight or branched alkyl chain with 1-10 C atom, cycloalkyl with 3-10 C atom, cycloalkylalkyl with 7-20 C atom, aralkyl with 7-20 C atom---wherein the aryl atomic group can be replaced by alkyl or halogen atom alternatively, phenyl---replaced by alkyl or halogen atom alternatively, or heterocycle---replaced by alkyl or halogen atom alternatively

R2 is group-CH (R21) R22, and wherein R21 is hydrogen atom, C ₁-C ₁₀-alkyl or optional substituted benzyl ring, R22 are optional substituted benzyl ring or naphthyl ring,

Or group-CH ₂CH (R23) R24, wherein R23 and R24 represent optional substituted benzyl ring,

R3 and R4 can be hydrogen atom or the alkyl with 1-10 C atom independently of one another, and R3 can also be a halogen atom,

R5 is the group that connects via atomic group Z

Wherein G be-C=C-,-C=N-,-N=C-, oxygen or sulphur atom, Z be direct key, oxygen or sulphur atom, group CH-R52 or-wherein R52 and R53 have the implication of hydrogen atom or alkyl to CHR52-CH-R53--independently of one another, n is that wherein R52 and R53 have the implication of hydrogen atom or alkyl to numeral 1 and 2---,-C ≡ C-three key or have E or the group-CR52=CR53-of Z configuration or C=CR52R53---independently of one another, and L is CH ₂Or the NH group, Q be carbonyl or-SO _xGroup, x=0,1 or 2 wherein, R51 is amino---alternatively by alkyl replace, straight or branched alkyl---replaced by halogen atom, hydroxyl or alkoxyl group alternatively or have the cycloalkyl of 3-7 ring members---is alternatively by halogen atom, hydroxyl or alkoxyl group replacement

R6 is group CH ₂-N (R61) R62, wherein R61 is hydrogen atom or alkyl independently of one another, R62 is the aralkyl or the heteroaralkyl of alkyl or the optional substituted 7-20 of having a C atom,

It can be group

The arbitrary orientation of described structure and all steric isomers, and with physiology on the acid that can tolerate or the salt of alkali.

In formula (1) compound, for instance,

Among the R1, the straight or branched alkyl chain: expression methyl, ethyl, n-propyl, sec.-propyl, just-, different-, tert-butyl, just-amyl group, 2,2-dimethyl propyl or 3-methyl butyl, just-hexyl, just-heptyl, just-octyl group, just-nonyl, just-decyl.Methyl or ethyl are preferred.

Phenyl is replaced by alkyl or halogen atom alternatively: expression phenyl, neighbour-,-, right-methyl-, ethyl-, propyl group-, isopropyl phenyl, 2,3-, 2,4-, 2,5-, 2,6-, 3,4-, 3, the 5-dimethyl-or-diethyl phenyl, neighbour-,-, right-fluoro-, chloro-, bromo-, iodophenyl, 2,3-, 2,4-, 2,5-, 2,6-, 3,4-, 3,5-two fluoro-, two chloro-, two bromo-or diiodo-phenyl or naphthyl.Phenyl is preferred.

Optional substituted furans or thienyl: represent unsubstituted 2-or 3-thienyl, 2-or 3-furyl or 3-methyl-, the 3-ethyl-, the 3-fluoro-, the 3-chloro-, the 3-bromo-, 3-iodo-2-furyl or-the 2-thienyl, the 4-methyl-, the 4-ethyl-, the 4-fluoro-, the 4-chloro-, the 4-bromo-, 4-iodo-2-furyl or-the 2-thienyl, the 5-methyl-, the 5-ethyl-, the 5-fluoro-, the 5-chloro-, the 5-bromo-, 5-iodo-2-furyl or-the 2-thienyl, the 2-methyl-, the 2-ethyl-, the 2-fluoro-, the 2-chloro-, the 2-bromo-, 2-iodo-3-furyl or-the 3-thienyl, the 4-methyl-, the 4-ethyl-, the 4-fluoro-, the 4-chloro-, the 4-bromo-, 4-iodo-3-furyl or-the 3-thienyl, the 5-methyl-, the 5-ethyl-, the 5-fluoro-, the 5-chloro-, the 5-bromo-, 5-iodo-3-furyl or-the 3-thienyl.2-thienyl or 2-furyl are preferred.

Aralkyl with 7-20 C atom: expression benzyl, the 1-phenylethyl,-propyl group,-butyl,-hexyl,-2-methylethyl,-2-ethyl diethyldithiocarbamate,-2, the 2-dimethyl ethyl, adjacent-, between-, right-methyl-, ethyl-, propyl group-, isopropyl benzyl, 2 ', 3 '-, 2 ', 4 '-, 2 ', 5 '-, 2 ', 6 '-, 3 ', 4 '-, 3 ', 5 '-dimethyl-or-the diethyl benzyl, 2 '-, 3 '-, 4 '-fluoro-, chloro-, bromo-, the iodine benzyl, 2 ', 3 '-, 2 ', 4 '-, 2 ', 5 '-, 2 ', 6 '-, 3 ', 4 '-, 3 ', 5 '-two fluoro-, two chloro-, two bromo-or diiodo-benzyl, 2-or 3-naphthyl methyl, the 2-phenylethyl, the 3-phenyl propyl, the 4-phenyl butyl, the 5-phenylpentyl.

C ₁-C ₆-alkyl: expression has the straight or branched alkyl of 1-6 C atom, for example methyl, ethyl, just-propyl group, sec.-propyl, just-, different-, tert-butyl, just-amyl group, 2,2-dimethyl propyl or 3-methyl butyl.

Cycloalkyl atomic group: representative ring propane, tetramethylene, pentamethylene, hexanaphthene, suberane, perhydronaphthalene atomic group.

The cycloalkylalkyl atomic group: the representative ring propyl group-, cyclobutyl-, cyclopentyl-, cyclohexyl-, suberyl methyl groups, 1-cyclopropyl-, the 1-cyclobutyl-, the 1-cyclopentyl-, the 1-cyclohexyl-, 1-suberyl ethyl atomic group, 2-cyclopropyl-, the 2-cyclobutyl-, the 2-cyclopentyl-, the 2-cyclohexyl-, 2-suberyl ethyl atomic group.

Heterocycle: represent unsubstituted 2-or 3-thienyl or 2-or 3-furyl or 3-methyl-, the 3-ethyl-, the 3-fluoro-, the 3-chloro-, the 3-bromo-, 3-iodo-2-furyl or-the 2-thienyl, the 4-methyl-, the 4-ethyl-, the 4-fluoro-, the 4-chloro-, the 4-bromo-, 4-iodo-2-furyl or-the 2-thienyl, the 5-methyl-, the 5-ethyl-, the 3-fluoro-, the 5-chloro-, the 5-bromo-, 5-iodo-2-furyl or-the 2-thienyl, the 2-methyl-, the 2-ethyl-, the 2-fluoro-, the 2-chloro-, the 2-bromo-, 2-iodo-3-furyl or-the 3-thienyl, the 4-methyl-, the 4-ethyl-, the 4-fluoro-, the 4-chloro-, the 4-bromo-, 4-iodo-3-furyl or-the 3-thienyl, the 5-methyl-, the 5-ethyl-, the 5-fluoro-, the 5-chloro-, the 5-bromo-, 5-iodo-3-furyl or-the 3-thienyl, unsubstituted 2-, 3-or 4-pyridyl, the 3-methyl-, the 3-ethyl-, the 3-fluoro-, the 3-chloro-, the 3-bromo-, 3-iodo-2-pyridyl, the 4-methyl-, the 4-ethyl-, the 4-fluoro-, the 4-chloro-, the 4-bromo-, 4-iodo-2-pyridyl, the 5-methyl-, the 5-ethyl-, the 5-fluoro-, the 5-chloro-, the 5-bromo-, 5-iodo-2-pyridyl, the 2-methyl-, the 2-ethyl-, the 2-fluoro-, the 2-chloro-, the 2-bromo-, 2-iodo-3-pyridyl, the 4-methyl-, the 4-ethyl-, the 4-fluoro-, the 4-chloro-, the 4-bromo-, 4-iodo-3-pyridyl, the 5-methyl-, the 5-ethyl-, the 5-fluoro-, the 5-chloro-, the 5-bromo-, 5-iodo-3-pyridyl, 2-, 4-, 5-, the 6-pyrimidyl, 3-, 4-, 5-, the 6-pyridazinyl, 2-or 3-pyrazinyl.

Among the R2, alkyl: expression has the straight or branched alkyl of 1-6 C atom, for example methyl, ethyl, just-propyl group, sec.-propyl, just-, different-, tert-butyl, just-amyl group, 2,2-dimethyl propyl or 3-methyl butyl.Hydrogen atom is preferred.

Optional substituted benzyl ring or naphthyl ring: expression phenyl, adjacent-, between-, right-methyl-,-ethyl-,-propyl group-,-isopropyl phenyl, 2,3-, 2,4-, 2,5-, 2,6-, 3,4-, 3, the 5-dimethyl-or-the diethyl phenyl, adjacent-, between-, right-fluoro-, chloro-, bromo-, iodophenyl, 2,3-, 2,4-, 2,5-, 2,6-, 3,4-, 3,5-two fluoro-, two chloro-, two bromo-or diiodo-phenyl, adjacent-, between-, right-the trihalogenmethyl phenyl, 2,3-, 2,4-, 2,5-, 2,6-, 3,4-, 3,5-two trihalogenmethyl phenyl, adjacent-, between-, right-methoxyl group-,-oxyethyl group-,-propoxy--,-isopropyl phenyl or naphthyl.2, the 5-difluorophenyl is preferred.

Among R3 and the R4, alkyl: expression has the straight or branched alkyl of 1-6 C atom, for example methyl, ethyl, just-propyl group, sec.-propyl, just-, different-, tert-butyl, just-amyl group, 2,2-dimethyl propyl or 3-methyl butyl.Hydrogen atom is preferred.

Among the R5, alkyl: expression has the straight or branched alkyl of 1-6 C atom, for example methyl, ethyl, just-propyl group, sec.-propyl, just-, different-, tert-butyl, just-amyl group, 2,2-dimethyl propyl or 3-methyl butyl.Hydrogen atom is preferred.

Among the R6, alkyl: expression has the straight or branched alkyl of 1-6 C atom, for example methyl, ethyl, just-propyl group, sec.-propyl, just-, different-, tert-butyl, just-amyl group, 2,2-dimethyl propyl or 3-methyl butyl.Methyl is preferred.

Heteroaralkyl with 7-20 C atom: expression 2-, 3-or 4-picolyl ,-ethyl or-propyl group, 2-or 3-furylmethyl ,-ethyl or-propyl group, 2-or 3-thenyl ,-ethyl or-propyl group, 2-, 3-, 4-, 5-, 6-or 7-indole methyl ,-ethyl or-propyl group.Benzyl is preferred.

Preferred formula (1) compound is such, wherein

It is group

Or

If R1 is group-CO-R11, R11 for example has preferred implication methyl, ethyl, sec.-propyl, phenyl, 2-thienyl and 2-furyl so.If R1 has-implication of CO-OR12, and R12 for example can be preferably methyl, ethyl or sec.-propyl so.

Preferred in addition such compound, wherein R2 is an aromatic group,---definitely being fluorine atom---by one or more halogen atoms on aromatic ring replaces, for example benzyl, for example 2 ', 6 '-difluorobenzyl.Same preferred such compound, wherein R3 and R4 have one at least---definitely be two all---is hydrogen atom.

The preferred meaning of Z is direct key or Sauerstoffatom, and G is preferably-the C=C-group.L is preferably the NH group, and Q is preferably carbonyl, and R51 is C ₁-C ₆-alkyl.Special preferred meaning about R61 is hydrogen atom or C ₁-C ₃-alkyl is a methyl definitely, about R62, is aralkyl atomic group, for example benzyl.

Compound (1) preferably prepares like this:

(a) make general formula (2) compound

Wherein R7 is a leavings group, for example halogen atom or alkyl-, perfluoroalkyl-or aryl sulfonyl, every other atomic group has implication shown in the compound (1),

With the reaction of general formula (3) compound,

R8-N(R61)R62 (3)

Wherein R8 is hydrogen atom or atoms metal, for example lithium, sodium, potassium, caesium, calcium or barium atom, and R61 and R62 have implication shown in the compound (1),

(b) make general formula (4) compound

Wherein R9 is group-OSO ₂C _nF _2n+1, halogen atom, definitely be bromine or iodine atom or another kind of leavings group, every other atomic group has implication shown in the compound (1),

With the reaction of general formula (5) compound,

Wherein R10 is the group that contains metal, for example trialkyltin, halo magnesium; The group that contains nonmetal, for example boron, silicon etc., for example dialkoxy boron or dihydroxyl boron; Hydroxyl or sulfydryl are converted into metal-salt alternatively, for example lithium, sodium, potassium, caesium, calcium, barium, silver or mantoquita; Group-C ≡ C-R31 or have E or the group-CR52=CR53R31 of Z configuration or-CR31=CR52R53, wherein R31 is the group that contains metal or nonmetal, for example boron, silicon etc., for example trialkyltin, halo magnesium, dialkoxy boron or dihydroxyl boron, every other atomic group has implication shown in the compound (1)

Reaction is with or without the participation of catalyzer, for example organic derivative of copper, nickel, palladium, platinum or described metal;

(c), then make general formula (6) compound if the Y in the compound (1) is a nitrogen-atoms

Wherein R32 is hydrogen atom or atoms metal, for example lithium, sodium, potassium, caesium, calcium, barium, silver or copper atom, and every other atomic group has implication shown in the compound (1),

With the reaction of general formula (7) compound,

R33R2 (7)

Wherein R33 is a leavings group, for example halogen atom or alkyl-, perfluoroalkyl-or aryl sulfonyl, R2 has implication shown in the compound (1), perhaps

(d), then make general formula (8) compound if the W in the compound (1) is a nitrogen-atoms

Wherein R32 is hydrogen atom or atoms metal, for example lithium, potassium, caesium, calcium, barium, silver or copper atom, and every other atomic group has implication shown in the compound (1),

With the reaction of general formula (9) compound,

R33-R1 (9)

Wherein R33 is a leavings group, for example halogen atom or alkyl-, perfluoroalkyl-or aryl sulfonyl, R1 has implication shown in the compound (1).

Can for example be used for male contraception, hormonotherapy, low and sterile treatment, female contraception and the tumour control of female fertility as the antagonist of gonadotropin releasing hormone according to compound of the present invention (1).

In male contraception, compound according to the present invention causes spermatogenetic reduction.Preferably, take and the male hormone Combined Preparation, for example testosterone or testosterone derivative, for example testosterone ester.The administration of testosterone derivative for example can be undertaken by injection, for example intramuscular Drug Storage injection.

In hormonotherapy, for example also be used for the treatment of endometriosis, leiomyoma of uterus and fibroma uteri, can unite alternatively and adopt compound (1) and other hormones, for example oestrogenic hormon is or/and progestogen.Combination according to GnRH antagonist of the present invention and tissue selectivity part estrogen agonist, for example raloxifene  is particularly preferred.And, can be used in the raising female fertility according to compound of the present invention (1), for example induced ovulation is used for the treatment of infertility.

On the other hand, compound (1) also is suitable for contraception of woman.Thereby, can be the 1st to 15 day of cycle with the GnRH antagonist with the oestrogenic hormon administration, preferred low-down estrogen dosage.The the 16th to 21 day of medicining cycle, add progestogen to the combination of oestrogenic hormon-GnRH antagonist.The GnRH antagonist can continue administration and go through the complete cycle.In this manner, can realize the reduction of hormone dosage and the reduction of non-physiology hormonal readiness side effect.In addition, the women of---for example acne, stearrhea and hirsutism---also can realize beneficial effect to suffering from polycystic ovarian syndrome and male hormone dependence disease.Compared with former medication, expection also improves periodic Control.Further indication is sexual gland protection, control ovarian stimulation/artificial reproductive technology, early stage infant development dysfunction, for example puberty precocity and the polycystic ovary of benign prostatic hyperplasia, chemotherapeutic period.

At last, GnRH agonist according to the present invention can also be used for the treatment of hormone-dependent tumor disease, and for example mammary cancer, prostate cancer, ovarian cancer and carcinoma of endometrium before the menopause are because their suppress endogenous sex steroid hormone.

Be suitable as the GnRH antagonist to people's administration according to compound of the present invention (1), but also be used for animal doctor's purpose, for example domestic and productivity animal and wildlife.

Administration can be carried out according to known way, for example oral, local, rectum, intravaginal, nose or injection.Oral administration is preferred.With compound (1) make can administration form, alternatively with pharmaceutically acceptable carrier and/or mixing diluents.Oral administration for example can carry out with solid form, and for example tablet, capsule, coating tablet or pulvis also can be the forms of drinkable solution still.Non-oral administration for example can carry out by intravenously, subcutaneous or intramuscularly or by ointment, creme or suppository.Alternatively, also might carry out administration with the slowly-releasing form.Dosage can be different because of indication type, disease seriousness, curee's age, sex, body weight and susceptibility.Preferably, dosage be 0.01 to 30mg, preferred especially 0.1 to 3mg, most preferably 0.1 to 1mg every kg body weight every day.Administration can be undertaken by single dose or some divided doses.

Enumerate some particularly preferred compounds (1) below:

Particularly preferred compound

The following example will further be set forth the present invention.

Embodiment

Embodiment 1

Ethyl 6-(4-kharophen phenoxy group)-5-(N-benzyl-N-methylamino-methyl)-1-(2 ', 6 '-difluorobenzyl)-1,4-dihydro-4-Oxoquinoline-3-carboxylic acid ester

With 78mg ethyl 6-(4-kharophen phenoxy group)-5-(chloromethyl)-1-(2 ', 6 '-difluorobenzyl)-1,4-dihydro-4-Oxoquinoline-3-carboxylic acid ester is dissolved in the 3ml dimethyl formamide, following at 0 ℃ with 84 μ l N-methylbenzylamines and 84 μ l N, the N-diisopropylethylamine is handled, and mixture was at room temperature stirred 20 hours.After adding the 20ml sodium bicarbonate aqueous solution, suction leaches precipitation, washes with water, is just using-hexane wash again, and is dry in the room temperature vacuum.Obtain the 70mg title compound.

NMR：δ＝1.3(t；3H；CH ₃)；1.9(s；3H；NCH ₃)；2.05(s；

3H；CH ₃)；3.55(s；2H；NCH ₂)；4.27(q；2H；

OCH ₂)；4.91(s；2H；NCH ₂)；5.68(s；2H；NCH ₂)；

6.85(d；2H；ArCH)；7.1-7.22(m；7H；ArCH)；

7.26(d；1H；ArCH)；?7.42-7.6(m；4H；ArCH)；

8.72(s；1H；NCH)；9.9(s；1H；NH)

MS：FAB：M +1＝526[M＝525]

Raw material ethyl 6-(4-kharophen phenoxy group)-5-(chloromethyl)-1-(2 ', 6 '-difluorobenzyl)-1,4-dihydro-4-Oxoquinoline-3-carboxylic acid ester obtains according to following manner.

A. ethyl 1,4-dihydro-6-fluoro-5-nitro-4-Oxoquinoline-3-carboxylic acid ester

50g 4-fluoro-3-N-methyl-p-nitroaniline and 69g ethoxy methylene diethyl malonate were heated 3 hours down at 120 ℃.Mixture is just being joined-hexane in, stirred 2 hours, suction leaches crystallisate, and is dry in the room temperature vacuum.Obtain 93g N-(4-fluoro-3-nitrophenyl) aminomethylene diethyl malonate.Be divided into 3 parts of 31g, join separately be heated in advance 260 ℃, 26.5% phenylbenzene and 73.5% phenyl ether (DOWTHERM A ^) the 150ml mixture in, mixture was stirred 30 minutes under this temperature.After the cooling, with 500ml just-hexane dilution, suction leaches precipitation.Obtain amounting to the ethyl 1 of 64g, 4-dihydro-6-fluoro-5-nitro-4-Oxoquinoline-3-carboxylic acid ester and 1, the mixture of 4-dihydro-6-fluoro-7-nitro-4-Oxoquinoline-3-carboxylic acid ester.

NMR：δ＝1.31(t；3H；CH ₃)；4.25(q；2H；OCH ₂)；7.88

(s；1H；ArCH；isomer?A)；7.92(s；1H；ArCH；

isomer?A)；8.1(d；1H；ArCH；isomer?B)；8.45

(d；1H；ArCH；isomer?B)；8.59+8.69(2s；

each?1H；NCH；A+B)

MS：EI：M ＝280[M＝280]

B. ethyl 1-(2 ', 6 '-difluorobenzyl)-1,4-dihydro-6-fluoro-5-nitro-4-Oxoquinoline-3-carboxylic acid ester

At room temperature, with 22g said mixture and 16g salt of wormwood and 23.7g 2, the 6-difluoro benzyl bromide stirred 5 hours in the 500ml dimethyl formamide.Reaction mixture is joined in the 1L aqueous ammonium chloride solution, use ethyl acetate extraction three times.Use dried over sodium sulfate, evaporation in a vacuum, add then 500ml just-hexane, mixture was stirred 15 minutes.Drain hexane mutually after, make resistates recrystallization from ethyl acetate.Obtain the 10.4g title compound.

NMR：δ＝1.3(t；3H；CH ₃)；4.25(q；2H；OCH ₂)；5.84(s；

2H；NCH ₂)；7.15-7.25(m；2H；ArCH)；7.45-

7.55(m；1H；ArCH)；7.92(dd；1H；ArCH)；8.04

-8.14(m；1H；ArCH)；8.96(s；1H；NCH)；

MS：EI：M ＝406[M＝406]

C. ethyl 6-(4-kharophen phenoxy group)-1-(2 ', 6 '-difluorobenzyl)-1,4-dihydro-5-nitro-4-Oxoquinoline-3-carboxylic acid ester

3.74g 4-acetoamidophenol is handled with 733mg sodium hydride (80%, mineral oil) in the 40ml dimethyl formamide, mixture was at room temperature stirred 15 minutes.This solution is joined in the 40ml dimethyl formamide solution of 5g above-claimed cpd then.After at room temperature 5 hours, mixture is joined in the frozen water, suction leaches precipitation.Through (eluent is methylene dichloride/2-propyl alcohol 95: 5) behind the silica gel chromatography purifying, obtain the 5.17g title compound.

NMR：δ＝1.3(t；3H；CH ₃)；2.03(s；3H；CH ₃)；4.27(q；

2H：OCH ₂)；5.79(s；2H；NCH ₂)；7.01(d；2H；

ArCH)；7.13-7.23(m；2H；ArCH)；7.43-7.56

(m；2H；ArCH)；7.61(d；2H；ArCH)；7.8(d；

1H；ArCH)；8.93(s；1H；NCH)；9.97(1H；S；

NCH)

MS：EI：M ＝537[M＝537]

D. ethyl 6-(4-kharophen phenoxy group)-5-amino-1-(2 ', 6 '-difluorobenzyl)-1,4-dihydro-4-Oxoquinoline-3-carboxylic acid ester

With 510mg palladium/carbon (10%) hydrogenation in 650ml methyl alcohol of 5.1g above-claimed cpd.Suction leaches catalyzer, and evaporation obtains the 4.55g title compound.

NMR：δ＝1.29(t；3H；CH ₃)；2.02(s；3H；CH ₃)；3.25(s；

2H；NH ₂)；4.25(q；2H；OCH ₂)；5.55(s；2H；

NCH ₂)；6.55(d；1H；ArCH)；6.86(d；2H；

ArCH)；7.07(d；1H；ArCH)；7.1-7.22(m；2H；

ArCH)；7.4-7.55(m；3H；ArCH)；8.71(s；1H；

NCH)；9.8(s；1H；NH)；

MS：EI：M ＝507[M＝507]

E. ethyl 6-(4-kharophen phenoxy group)-1-(2 ', 6 '-difluorobenzyl)-1,4-dihydro-5-iodo-4-Oxoquinoline-3-carboxylic acid ester

The 2g above-claimed cpd is dissolved in the mixture of the 24ml vitriol oil and 12ml water, under 0 ℃, mixture is handled with the 4ml aqueous solution of 325mg Sodium Nitrite.After 15 minutes, mixture is transferred to pH3 with sodium hydrogen carbonate solution, add the 100mg urea.The 0.5ml aqueous solution that adds the 723mg potassiumiodide then at room temperature stirred mixture one hour.With methylene chloride (95: 5, v/v) extraction after, organic phase is washed with sodium thiosulfate solution, drying, the evaporation.Through behind the silica gel chromatography purifying (eluent is the methylene dichloride that contains the 0-15% Virahol), obtain the 697mg title compound.

NMR：δ＝1.3(t；3H；CH ₃)；2.02(s；3H；CH ₃)；4.25(q；

2H；OCH ₂)；5.71(s；2H；NCH ₂)；6.83(d；2H；

ArCH)；7.1-7.2(2H；m；ArCH)；7.3(d；1H；

ArCH)；7.41-7.63(m；4H；ArCH)；8.83(s；

1H；N-CH)；9.95(s；1H；NH)

MS：es：M +1＝493[M＝492]

F. ethyl 6-(4-kharophen phenoxy group)-1-(2 ', 6 '-difluorobenzyl)-1,4-dihydro-4-oxo-5-styryl quinoline-3-carboxylic acid ester

The above-mentioned iodo compound of 550mg, 198mg styryl were stirred 6 hours down at 80 ℃ for boric acid, 55mg 10 4 (triphenyl phosphine) palladium (0), 1.1ml 2 molar sodium carbonate solution, 2.2ml ethanol and 22ml toluene.Add entry then, the mixture dichloromethane extraction washs organic phase with sodium chloride solution, drying, evaporation.Resistates obtains the 902mg title compound through silica gel chromatography purifying (eluent is the methylene dichloride that contains the 0-10% Virahol).

NMR：δ＝1.28(t；3H；CH ₃)；2.0(s；3H；CH ₃)；4.24(q；

2H；OCH ₂)；5.7(s；2H；N-CH ₂)；6.75-6.85(m；

3H；ArCH；CH＝CH)；7.08-7.28(m；3H；ArCH)；

7.3-7.68(M；9H；ArCH)；7.86(d；1H；

CH＝CH)；8.75(s；1H；NCH)；9.72(1H；S；NH)

MS：es：M +1＝595[M＝594]

G. ethyl 6-(4-kharophen phenoxy group)-1-(2 ', 6 '-difluorobenzyl)-1,4-dihydro-5-formyl radical-4-Oxoquinoline-3-carboxylic acid ester

The above-mentioned compound of styryl of 450mg is dissolved in 25ml tetrahydrofuran (THF) and 7ml water, handles with the uncle-butanol solution of 0.11ml 2.5% perosmic anhydride.After at room temperature stirring 15 minutes, add 482mg, add 100mg after 20 and 22 hours, add the 100mg sodium periodate after 24 hours.After 26 hours,, use ethyl acetate extraction with the mixture dilute with water.With organic phase with dried over sodium sulfate after, the evaporation, obtain the 352mg title compound, be foam.

NMR：δ＝1.29(t；3H；CH ₃)；2.01(s；3H；CH ₃)；4.27(q；

2H；OCH ₂)；5.71(s；NCH ₂)；7.12-7.22(m；2H；

ArCH)；7.4(d；1H；ArCH)；7.43-7.64(m；3H；

ArCH)；7.72(d；1H；ArCH)；9.0(d；1H；NCH)；

9.97(s；1H；NH)；10.44(s；1H；CHO)

MS：FAB：M +1＝521[M＝520]

H. ethyl 6-(4-kharophen phenoxy group)-1-(2 ', 6 '-difluorobenzyl)-1,4-dihydro-5-methylol-4-Oxoquinoline-3-carboxylic acid ester

The above-mentioned aldehyde of 300mg is dissolved in 13.8ml acetate, handles, 10 minutes at interval with 3 parts of sodium borohydrides of each 10mg.Behind the dilute with water, the mixture ethyl acetate extraction is washed till neutrality with organic phase with sodium hydrogen carbonate solution.After dried over sodium sulfate, evaporation.Through behind the silica gel chromatography purifying (eluent is the methylene dichloride that contains the 0-10% Virahol), obtain the 129mg title compound, be foam.

NMR：δ＝1.31(t；3H；CH ₃)；2.02(s；3H；CH ₃)；4.27(q；

2H；OCH ₂)；4.86(d；2H；OCH ₂)；5.12(t；1H；

OH)；5.8(s；2H；NCH ₂)；6.85(d；2H；ArCH)；

7.12-7.22(m；2H；ArCH)；7.38(d；1H；

ArCH)；7.42-7.59(m；3H；ArCH)；7.63(d；

1H；ArCH)；8.94(s；1H；NCH)；9.5(s；1；NH)

MS：FAB：M +1＝523[M＝522]

I. ethyl 6-(4-kharophen phenoxy group)-5-chloromethyl-1-(2 ', 6 '-difluorobenzyl)-1,4-dihydro-4-Oxoquinoline-3-carboxylic acid ester

The above-mentioned alcohol of 80mg is dissolved in the 2ml methylene dichloride, handles with the 0.1ml thionyl chloride.After at room temperature 20 minutes, add entry, the mixture dichloromethane extraction, organic phase is washed with sodium chloride solution.After dried over sodium sulfate, evaporation.Obtain the 80mg title compound, be foam.

NMR：δ＝1.33(t；3H；CH ₃)；2.07(s；3H；CH ₃)；4.3(q；

2H；OCH ₂)；5.66(s；2H；CH ₂Cl)；5.75(s；2H；

NCH ₂)；6.98(d；2H；ArCH)；7.13-7.25(m；

2H；ArCH)；7.3(d；1H；ArCH)；7.53(t；1H；

ArCH)；7.61(d；2H；ArCH)；7.68(d；1H；

ArCH)；8.82(s；1H；NCH)；9.98(s；1H；NH)

MS：es：M +1＝541/543[M＝540/542]

Embodiment 2

2-propyl group 6-(4-kharophen phenoxy group)-5-(N-benzyl-N-methylamino-methyl)-1-(2 ', 6 '-difluorobenzyl)-1,4-dihydro-4-Oxoquinoline-3-carboxylic acid ester

With ethyl 6-(4-kharophen phenoxy group)-5-(N-benzyl-N-methylamino-methyl)-1-(2 ', 6 '-difluorobenzyl)-1,4-dihydro-4-Oxoquinoline-3-carboxylic acid ester and titanium tetraisopropylate heat in Virahol, generate this compound.

Embodiment 3

Ethyl 5-(N-benzyl-N-methylamino-methyl)-1-(2 ', 6 '-difluorobenzyl)-1,4-dihydro-6-(4-isobutyryl amino-benzene oxygen)-4-Oxoquinoline-3-carboxylic acid ester

Be similar to embodiment 1, from ethyl 5-(chloromethyl)-1-(2 ', 6 '-difluorobenzyl)-1,4-dihydro-6-(4-isobutyryl amino-benzene oxygen)-4-Oxoquinoline-3-carboxylic acid ester and N-methylbenzylamine generate this compound, are foam.

A. ethyl 5-(chloromethyl)-1-(2 ', 6 '-difluorobenzyl)-1,4-dihydro-6-(4-isobutyryl amino-benzene oxygen)-4-Oxoquinoline-3-carboxylic acid ester

In embodiment 1/c, replace the 4-acetoamidophenol, further be similar to embodiment 1/d to embodiment 1/i reaction, obtain this compound with 4-isobutyryl amino-phenol.

Embodiment 4

3-ethanoyl-6-(4-acetylamino phenyl)-1-benzyl-5-(N-benzyl-N-methylamino-methyl) phthalazines-4-ketone

In the presence of alkali, for example yellow soda ash or sodium hydroxide make 6-(4-acetylamino phenyl)-1-benzyl-5-(N-benzyl-N-methylamino-methyl) phthalazines-4-ketone and Acetyl Chloride 98Min. or acetic anhydride, obtain title compound.

6-(4-acetylamino phenyl)-1-benzyl-5-(N-benzyl-N-methylamino-methyl) phthalazines-4-ketone obtains according to following manner.

A.6-(4-acetylamino phenyl)-1-benzyl-5-opacin piperazine-4-ketone

Be similar to embodiment 1/f, make 1-benzyl-5,6-diiodo-phthalazines-4-ketone (Indian J.Chem.16B, 1978,301-304) with 1 equivalent 4-acetylamino phenyl for acid reaction.Through the silica gel chromatography purifying, obtain title compound.

B.6-(4-acetylamino phenyl)-1-benzyl-5-chloromethyl phthalazines-4-ketone

Be similar to embodiment 1/f-i 6-(4-acetylamino phenyl)-1-benzyl-5-opacin piperazine-4-ketone is further reacted, obtain title compound.

Embodiment 5

Ethyl 6-(4-kharophen phenoxy group)-5-(N-benzyl-N-methylamino-methyl)-1,4-dihydro-4-oxo-1-(2 '-trifluoromethyl benzyl) quinoline-3-carboxylic acid ester

With 130mg (0.226mmol) ethyl 6-(4-kharophen phenoxy group)-5-(chloromethyl)-1,4-dihydro-4-oxo-1-(2 '-trifluoromethyl benzyl) quinoline-3-carboxylic acid ester is dissolved in 5ml DMF, with 125 μ l N, N-diisopropylethylamine and 12 μ l (0.97mmol) N-benzyl methylamines are handled under-5 ℃.After being warmed to room temperature, reaction mixture was stirred 20 hours, join then in the 50ml saturated sodium bicarbonate solution.Suction leaches solid, washes with water, and is dry in a vacuum.Be further purified by silica gel chromatography, eluent is formed (R by 90 parts of methylene dichloride, 10 parts of ethanol and 1 part of dense ammonia _f: 0.38) .2.Obtain the 45mg title compound, be foam.

MS/ mole peak, M ⁺=658

Use 2 '-trifluoromethyl benzyl bromo for 2,6-difluoromethyl bromotoluene, be similar to the described approach of embodiment 1a to 1i and prepare raw material ethyl 6-(4-kharophen phenoxy group)-5-(chloromethyl)-1,4-dihydro-4-oxo-1-(2 '-trifluoromethyl benzyl) quinoline-3-carboxylic acid ester.

Embodiment 6

Ethyl 6-(4-amino-carbonyl phenoxy group)-5-(N-benzyl-N-methylamino-methyl)-1-(2 ', 6 '-difluorobenzyl)-1,4-dihydro-4-Oxoquinoline-3-carboxylic acid ester

With 35mg (0.061mmol) ethyl 6-(4-amino-carbonyl phenoxy group)-5-(chloromethyl)-1-(2 ', 6 '-difluorobenzyl)-1,4-dihydro-4-Oxoquinoline-3-carboxylic acid ester is dissolved in 1.3 μ l DMF, with 35 μ l N, N-diisopropylethylamine and 35 μ l (0.25mmol) N-methylbenzylamines are handled under 0 ℃.After being warmed to room temperature, reaction mixture was stirred 20 hours, join then in the 10ml saturated sodium bicarbonate solution.Suction leaches the solid that is generated, and water and hexane wash are in a vacuum through five phosphorus oxide dryings.Obtain the 27mg title compound, be foam.

MS(esi)：M ⁺+1＝626(M＝625)

Use 4-hydroxy-n-methyl benzamide to replace the 4-acetoamidophenol, be similar to the described approach of embodiment 1a to 1i and prepare raw material ethyl 6-(4-amino-carbonyl phenoxy group)-5-(chloromethyl)-1-(2 ', 6 '-difluorobenzyl)-1,4-dihydro-4-Oxoquinoline-3-carboxylic acid ester.

Embodiment 7

Ethyl 6-(4-kharophen phenoxy group)-5-(N-benzyl-N-methylamino-methyl)-1-(1-menaphthyl)-1,4-dihydro-4-Oxoquinoline-3-carboxylic acid ester

With 45mg (0.081mmol) ethyl 6-(4-kharophen phenoxy group)-5-(chloromethyl)-1-(1-menaphthyl)-1,4-dihydro-4-Oxoquinoline-3-carboxylic acid ester is dissolved in 1.7ml DMF, with 46 μ l N, N-diisopropylethylamine and 46 μ l (0.33mmol) N-methylbenzylamines are handled under 0 ℃.After being warmed to room temperature, reaction mixture was stirred 20 hours, join then in the 15ml saturated sodium bicarbonate solution.Suction leaches the solid that is generated, and water and hexane wash are in a vacuum through five phosphorus oxide dryings.Obtain the 32mg title compound, be foam.

MS(esi)：M ⁺+1＝639(M＝638)

Use 1 chloromethyl naphthalene to replace 2,6-difluoromethyl bromotoluene is similar to the described approach of embodiment 1a to 1i and prepares raw material ethyl 6-(4-kharophen phenoxy group)-5-(chloromethyl)-1-(menaphthyl)-1,4-dihydro-4-Oxoquinoline-3-carboxylic acid ester.

Embodiment 8

The proof of antagonistic action

A) material

Buserelin is that (Frankfurt/Main Germany) obtains from Welding.Utilize chloramine-t method, compound is used ¹²⁵I and Na ¹²⁵I mark (4000Ci/mmol; Amersham-Buchler, Braunschweig, Germany).Material behind the mark is gone up through the reversed-phase HPLC purifying, with 50% acetonitrile/0.15% trifluoroacetic acid wash-out, flow velocity 0.5ml/min at Spherisorb ODS II post (250 * 4mm, particle diameter 3 μ m).Specific activity is 2000Ci/mmol.

Every other chemical all obtains from commercial source, and purity is the available superlative degree.

B) cell cultures

(Nunc on 245 * 245 * 20mm), is containing penicillin (100I.U./ml), Streptomycin sulphate (0.1mg/ml), glutamine (0.01mol/l) and 10% foetal calf serum (FCS in the plastics tissue culture plate; PAA Laboratories, Coelbe, DMEM substratum (Gibco-BRL Germany), Eggenstein-Leopoldshafen, Germany) (Bilezikjian et al., Mol.Endocrinol 5 (1991), 347-355) to cultivate α T3-1 cell in.Except using Ham ' sF12 substratum (Gibco-BRL), cultivate under the same conditions the CHO-3 cell (Schmidet al., J.Biol.Chem.275 (2000), 9193-9200).

10 fused cell culture plates are washed twice with 50ml phosphate buffered salt solution (PBS).Scrape with the rubber scraper and to get, cell harvesting in 5ml PBS, with the centrifugal 10min of 800rpm, is carried out sedimentation in laboratory centrifuge (Heraeus).The granular precipitation of cell is suspended among 5ml 0.25mol/l sucrose/0.01mol/l trolamine pH 7.4 once more.Through three cycles freezing in dry ice/ethanol bath, that at room temperature melt, make cytolysis.Lysate with the centrifugal 10min of 900rpm, is discarded sediment.With supernatant liquor in Sorvall SS34 whirler with 18, the centrifugal 30min of 000rpm.Granular precipitation (cytolemma) is suspended in 5ml measures in the damping fluid (0.25mol/l sucrose, 0.01mol/l trolamine pH 7.5,1mg/ml Protalbinic acid), per 200 μ l are stored under-20 ℃.(Anal.Biochem.72 (1976), method 248-254) is carried out protein determination according to Bradford.

C. receptor determination

Carry out combination research about competition curve, triplicate.Sample contains 60 μ l cytolemma suspensions (about α T3-1 cell is 10 μ g protein, is 40 μ g protein about the CHO-3 cell), 20 μ l ¹²⁵The buserelin of I mark (about competition curve is 100, the every increment of 000Ipm this, about saturation experiments 1500 and 200, between the 000Ipm) and 20 μ l confession try damping fluid or test compound solution.Test compound is dissolved in distilled water or 50% ethanol.In for the examination damping fluid, prepare serial dilutions (5 * 10 ^-6Mol/l to 5 * 10 ^-12Mol/l).At excessive unmarked buserelin (10 ^-6Mol/l) non-specific binding is measured in existence down.Sample is at room temperature cultivated 30min.Utilize Amicon 10x collecting device to filter (Whatman GF/C filter, diameter 2.5cm) and separate bonded and free part, with 5ml 0.02mol/l tris/HCl pH7.4 washed twice.With filter with 0.3% polymine (Serva; Heidelberg, Germany) moistening 30min, purpose is to reduce non-specific binding.In 5 passage gamma counters (Wallac LKB1470Wizard), measure the radioactivity that filter kept.

Claims

1, general formula (1) compound:

Wherein

R2 is group-CH (R21) R22, and wherein R21 is hydrogen atom, C ₁-C ₁₀-alkyl or optional substituted benzyl ring, R22 are optional substituted benzyl ring or naphthyl ring, or group-CH ₂CH (R23) R24, wherein R23 and R24 represent optional substituted benzyl ring,

R5 is the group that connects via atomic group Z

Wherein G be-C=C-,-C=N-,-N=C-, oxygen or sulphur atom, Z be direct key, oxygen or sulphur atom, group CH-R52 or-CHR52-CH-R53----wherein R52 and R53 have the implication of hydrogen atom or alkyl independently of one another, n is that wherein R52 and R53 have the implication of hydrogen atom or alkyl to numeral 1 and 2---,-C ≡ C-three key or have E or the group-CR52=CR53-of Z configuration or C=CR52R53---independently of one another, and L is CH ₂Or the NH group, Q be carbonyl or-SO _xGroup, x=0,1 or 2 wherein, R51 is amino---alternatively by alkyl replace, straight or branched alkyl---replaced by halogen atom, hydroxyl or alkoxyl group alternatively or have the cycloalkyl of 3-7 ring members---is alternatively by halogen atom, hydroxyl or alkoxyl group replacement

It can be group

2, as claim 1 claimed compounds, it is characterized in that It is group

Or

3,, it is characterized in that R1 is group-CO-R11 as claim 1 or 2 claimed compounds.

4,, it is characterized in that R11 is selected from methyl, ethyl, sec.-propyl, phenyl, 2-thienyl and 2-furyl as claim 3 claimed compounds.

5,, it is characterized in that R1 is group-CO-OR12 as claim 1 or 2 claimed compounds.

6,, it is characterized in that R12 is selected from methyl, ethyl or sec.-propyl as claim 5 claimed compounds.

7,, it is characterized in that R2 is 2 ', 5 '-difluorobenzyl as one of claim 1 to 6 claimed compounds.

8,, it is characterized in that R3 and R4 are hydrogen atoms as one of claim 1 to 7 claimed compounds.

9,, it is characterized in that Z is direct key or Sauerstoffatom as one of claim 1 to 8 claimed compounds.

10,, it is characterized in that G is-C=C-as one of claim 1 to 9 claimed compounds.

11,, it is characterized in that L is the NH group as one of claim 1 to 10 claimed compounds.

12, as one of claim 1 to 11 claimed compounds, it is characterized in that Q is a carbonyl, R51 is C ₁-C ₆-alkyl.

13,, it is characterized in that R61 is hydrogen atom or methyl, or/and R62 is a benzyl as one of claim 1 to 12 claimed compounds.

14, as the purposes of one of claim 1 to 13 claimed compounds as gonadotropin releasing hormone (GnRH) antagonist.

15,, be used for male contraception, hormonotherapy, low and sterile treatment, female contraception and the tumour control of female fertility as claim 14 purposes required for protection.

16, the preparation method of general formula (1) compound, this method is

(a) make general formula (2) compound

Wherein R7 is a leavings group, and every other atomic group has implication shown in the compound (1),

With the reaction of general formula (3) compound,

R8-N(R61)R62 (3)

Wherein R8 is hydrogen atom or atoms metal, and R61 and R62 have implication shown in the compound (1),

(b) make general formula (4) compound

With the reaction of general formula (5) compound,

Wherein R10 contains metal or nonmetallic group, hydroxyl or sulfydryl---be converted into metal-salt, group-C ≡ C-R31 alternatively have E or the group-CR52=CR53R31 of Z configuration or-CR31=CR52R53, wherein R31 contains metal or nonmetallic group, every other atomic group has implication shown in the compound (1)

Reaction is with or without the participation of catalyzer;

Wherein R32 is hydrogen atom or atoms metal, and every other atomic group has implication shown in the compound (1),

With the reaction of general formula (7) compound,

R33R2 (7)

Wherein R33 is a leavings group, and R2 has implication shown in the compound (1), perhaps

With the reaction of general formula (9) compound,

R33-R1 (9)

Wherein R33 is a leavings group, and R1 has implication shown in the compound (1).