[go: up one dir, main page]

WO1997004778A1 - Composes pyridiniques, intermediaires, processus industriels, compositions et procedes - Google Patents

Composes pyridiniques, intermediaires, processus industriels, compositions et procedes Download PDF

Info

Publication number
WO1997004778A1
WO1997004778A1 PCT/US1996/011958 US9611958W WO9704778A1 WO 1997004778 A1 WO1997004778 A1 WO 1997004778A1 US 9611958 W US9611958 W US 9611958W WO 9704778 A1 WO9704778 A1 WO 9704778A1
Authority
WO
WIPO (PCT)
Prior art keywords
arh
compound
phenyl
loweralkyl
benzoyloxy
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/US1996/011958
Other languages
English (en)
Inventor
Henry U. Bryant
Don R. Finley
Ken Matsumoto
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Eli Lilly and Co
Original Assignee
Eli Lilly and Co
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Eli Lilly and Co filed Critical Eli Lilly and Co
Priority to JP9507648A priority Critical patent/JPH11510163A/ja
Priority to AU65030/96A priority patent/AU6503096A/en
Publication of WO1997004778A1 publication Critical patent/WO1997004778A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/24—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with substituted hydrocarbon radicals attached to ring carbon atoms
    • C07D213/28—Radicals substituted by singly-bound oxygen or sulphur atoms
    • C07D213/30—Oxygen atoms
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4418—Non condensed pyridines; Hydrogenated derivatives thereof having a carbocyclic group directly attached to the heterocyclic ring, e.g. cyproheptadine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
    • A61K31/4439—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. omeprazole
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
    • A61K31/444—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a six-membered ring with nitrogen as a ring heteroatom, e.g. amrinone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/06—Antihyperlipidemics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P5/00—Drugs for disorders of the endocrine system
    • A61P5/24—Drugs for disorders of the endocrine system of the sex hormones
    • A61P5/32—Antioestrogens

Definitions

  • This invention relates to the fields of pharmaceutical and organic chemistry and provides novel pyridine compounds which are useful for the treatment of the various medical indications associated with post-menopausal syndrome.
  • the present invention further relates to intermediate compounds and processes useful for preparing the pharmaceutically active compounds of the present invention, and pharmaceutical compositions.
  • Post-menopausal syndrome is a term used to describe various pathological conditions which frequently affect women who have entered into or completed the physiological metamorphosis known as menopause. Although numerous pathologies are contemplated by the use of this term, three major effects of post-menopausal syndrome are the source of the greatest long-term medical concern: osteoporosis, cardiovascular effects such as hyperlipidaemia, and estrogen-dependent cancer, particularly breast and uterine cancer.
  • Osteoporosis describes a group of diseases which arise from diverse etiologies, but which are characterized by the net loss of bone mass per unit volume. The consequence of this loss of bone mass and resulting bone fracture is the failure of the skeleton to provide adequate structural support for the body.
  • One of the most common types of osteoporosis is that associated with menopause. Most women lose from about 20% to about 60% of the bone mass in the trabecular compartment of the bone within 3 to 6 years after the cessation of menses. This rapid loss is generally- associated with an increase of bone resorption and formation. However, the resorptive cycle is more dominant and the result is a net loss of bone mass. Osteoporosis is a common and serious disease among post-menopausal women.
  • osteoporosis is not generally thought of as a life threatening condition, a 20% to 30% mortality rate is related with hip fractures in elderly women. A large percentage of this mortality rate can be directly associated with post- menopausal osteoporosis.
  • the most vulnerable tissue in the bone to the effects of post-menopausal osteoporosis is the trabecular bone.
  • This tissue is often referred to as spongy or cancellous bone and is particularly concentrated near the ends of the bone (near the joints) and in the vertebrae of the spine.
  • the trabecular tissue is characterized by small osteoid structures which inter-connect with each other, as well as the more solid and dense cortical tissue which makes up the outer surface and central shaft of the bone. This inter-connected network of trabeculae gives lateral support to the outer cortical structure and is critical to the bio- mechanical strength of the overall structure.
  • post ⁇ menopausal osteoporosis In post ⁇ menopausal osteoporosis, it is, primarily, the net resorption and loss of the trabeculae which leads to the failure and fracture of bone.
  • the most common fractures are those associated with bones which are highly dependent on trabecular support, e.g., the vertebrae, the neck of the weight bearing bones such as the femur and the fore-arm.
  • hip fracture, collies fractures, and vertebral crush fractures are hall-marks of post-menopausal osteoporosis.
  • the only generally accepted method for treatment of post-menopausal osteoporosis is estrogen replacement therapy. Although therapy is generally successful, patient compliance with the therapy is low primarily because estrogen treatment frequently produces undesirable side effects.
  • estrogen replacement therapy It has been reported in the literature that post ⁇ menopausal women having estrogen replacement therapy have a return of serum lipid levels to concentrations to those of the pre-menopausal state. Thus, estrogen would appear to be a reasonable treatment for this condition. However, the side-effects of estrogen replacement therapy are not acceptable to many women, thus limiting the use of this therapy. An ideal therapy for this condition would be an agent which would regulate the serum lipid level as does estrogen, but would be devoid of the side-effects and risks associated with estrogen therapy.
  • the third major pathology associated with post ⁇ menopausal syndrome is estrogen-dependent breast cancer and, to a lesser extent, estrogen-dependent cancers of other organs, particularly the uterus.
  • neoplasms are not solely limited to a post-menopausal women, they are more prevalent in the older, post-menopausal population.
  • Current che otherapy of these cancers has relied heavily on the use of anti-estrogen compounds such as, for example, tamoxifen.
  • anti-estrogen compounds such as, for example, tamoxifen.
  • these agents may have stimulatory effects on certain cancer cell populations in the uterus due to their estrogenic (agonist) properties and they may, therefore, be contraproductive in some cases.
  • a better therapy for the treatment of these cancers would be an agent which is an anti-estrogen compound having negligible or no estrogen agonist properties on reproductive tissues.
  • the present invention provides new pyridine compounds, pharmaceutical compositions thereof, and methods of using such compounds for the treatment of post-menopausal syndrome and other estrogen-related pathological conditions.
  • vascular restenosis after percutaneous transluminal coronary angioplasty has been shown to be a tissue response characterized by an early and late phase.
  • the early phase occurring hours to days after PTCA is due to thrombosis with some vasospasms while the late phase appears to be dominated by excessive proliferation and migration of aortal smooth muscle cells.
  • the increased cell motility and colonization by such muscle cells and macrophages contribute significantly to the pathogenesis of the disease.
  • vascular aortal smooth muscle cells may be the primary mechanism to the reocclusion of coronary arteries following PTCA, atherectomy, laser angioplasty and arterial bypass graft surgery. See "Intimal Proliferation of Smooth Muscle Cells as an Explanation for Recurrent Coronary Artery Stenosis after Percutaneous Transluminal Coronary Angioplasty," Austin et al . , Journal of the American College of Cardiology. 8 : 369-375 (Aug. 1985).
  • vascular restenosis remains a major long term complication following surgical intervention of blocked arteries by percutaneous transluminal coronary angioplasty (PTCA) , atherectomy, laser angioplasty and arterial bypass graft surgery. In about 35% of the patients who undergo PTCA, reocclusion occurs within three to six months after the procedure.
  • the current strategies for treating vascular restenosis include mechanical intervention by devices such as stents or pharmacologic therapies including heparin, low molecular weight heparin, coumarin, aspirin, fish oil, calcium antagonist, steroids, and prostacyclin. These strategies have failed to curb the reocclusion rate and have been ineffective for the treatment and prevention of vascular restenosis.
  • the present invention provides for the use of compounds as smooth aortal muscle cell proliferation inhibitors and, thus inhibitors of restenosis.
  • the present invention is directed to compounds of Formula I:
  • n 2 or 3;
  • R is dimethylamino, diethylamino, 1-piperidinyl, 1- pyrrolidinyl, 4-morpholinyl, or 1-hexamethyleneimino;
  • the present invention is also directed to methods employing and compositions comprising these compounds, for the treatment of post menopausal syndrome.
  • the methods may additionally employ, and the compositions additionally comprise, an estrogen or progestin.
  • R 1 methyl
  • R 2 hydroxy
  • a particularly preferred compound is 2-[4-[2-(1-pyrrolidinyl) ethoxy]phenyl]-3-(4-hydroxyphenyl)-6-methylpyridine, including pharmaceutically acceptable salts.
  • Also provided by the present invention is a process for preparing a compound of formula la:
  • n 2 or 3;
  • R is dimethylamino, diethylamino, 1-piperidinyl, 1- pyrrolidinyl, 4-morpholinyl, or 1-hexamethyleneimino;
  • R la is hydrogen, loweralkyl of C- L -C 4 , phenyl, or mono- or disubstituted phenyl wherein each substituent is independently halo, methyl, Ci-Ce-alkoxy, benzyloxy, Ci-C ⁇ - alkanoyloxy, benzoyloxy, substituted benzoyloxy bearing 1 to 3 substituents each of which is independently halo, C 1 -C 4 - loweralkyl, or C ⁇ -C 4 ⁇ loweralkoxy, Ci-Cs-alkoxycarbonyloxy, or C 4 ⁇ C6-alkylsulfonyloxy;
  • R 2a is hydrogen, Ci-C6-alkoxy, benzyloxy, Ci-C ⁇ - alkanoyloxy, benzoyloxy, substituted benzoyloxy bearing 1 to 3 substituents each of which is independently halo, C1-C4- loweralkyl, or C ⁇ -C4-loweralkoxy
  • the compounds of the present invention are prepared by the following alkylation reaction:
  • Hydroxy protection is well known; see, for example, Protective Groups in Organic Chemistry, Plenum Press (London and New York, 1973); Protecting Groups in Organic Synthesis. Wiley (New York, 1981); and The Peptides, Vol. I, Schrooder and Lubke, Academic Press (London and New York, 1965) . Hydroxyl protection is also discussed in U.S. 4,418,068, which is incorporated herein by reference.
  • Suitable hydroxy protected groups are C ⁇ -C ⁇ alkoxy, benzyloxy, Ci-C ⁇ -alkanoyloxy, benzoyloxy, substituted benzoyloxy as previously defined, Ci-Cs-alkoxycarbonyloxy, and C 4 -C 6 ⁇ alkylsulfonyloxy.
  • the alkylation reaction is carried out by contacting the reactants, typically in an inert solvent. Suitable such solvents include methyl ethyl ketone, acetone, THF, dimethylformamide, and DMSO.
  • a base is supplied to serve as a hydrogen halide acceptor.
  • the reaction goes forward at temperatures over a wide range but is generally conducted at temperatures from 0 to 100 * C.
  • the reaction consumes the reactants and base in equimolar amounts; preferably, the reactants are supplied in equimolar amounts and the base is supplied in an excess amount. Isolation and preparation are carried out in standard procedures.
  • the free-base form of formula I compounds can be used in the methods of the present invention, it is often preferred to prepare and use a pharmaceutically acceptable salt form.
  • the compounds used in the methods of this invention primarily form pharmaceutically acceptable acid addition salts with a wide variety of organic and inorganic acids, and include the physiologically acceptable salts which are often used in pharmaceutical chemistry. Such salts are also part of this invention.
  • Typical inorganic acids used to form such salts include hydrochloric, hydrobromic, hydroiodic, nitric, sulfuric, phosphoric, hypophosphoric, and the like.
  • Salts derived from organic acids such as aliphatic mono and dicarboxylic acids, phenyl substituted alkanoic acids, hydroxyalkanoic and hydroxyalkandioic acids, aromatic acids, aliphatic and aromatic sulfonic acids, may also be used.
  • Such pharmaceutically acceptable salts thus include acetate, phenylacetate, trifluoroacetate, acrylate, ascorbate, benzoate, chlorobenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, methylbenzoate, o-acetoxybenzoate, naphthalene-2-benzoate, bromide, isobutyrate, phenylbutyrate, ⁇ -hydroxybutyrate, butyne-1,4-dioate, hexyne-1,4-dioate, caprate, caprylate, chloride, cinnamate, citrate, formate, fumarate, glycollate, heptanoate, hippurate, lactate, malate, maleate, hydroxymaleate, malonate, mandelate, mesylate, nicotinate, isonicotinate, nitrate, oxalate, phthalate, terephthalate, phosphate, monohydrogenphosphat
  • the pharmaceutically acceptable acid addition salts are typically formed by reacting a compound of formula I with an equimolar or excess amount of acid.
  • the reactants are generally combined in a mutual solvent such as diethyl ether or ethyl acetate.
  • the salt normally precipitates out of solution within about one hour to 10 days and can be isolated by filtration or the solvent can be stripped off by conventional means.
  • the pharmaceutically acceptable salts generally have enhanced solubility characteristics compared to the compound from which they are derived, and thus are often more amenable to formulation as liquids or emulsions.
  • Diethylamine hydrochloride (76.7 g, 70 mmol) was added portion wise at rt to a 200 mL aqueous solution of potassium cyanide (45.5 g, 70 mmol) with stirring and warmed to 60-65 °C.
  • a MeOH/THF (300 ml, 1:1) solution of 4- (benzyloxy) - benzaldehyde (100 g, 47 mmol) (Aldrich) was added in a thin stream to the aqueous solution and stirred for 16 hr at -62 °C. The reaction mixture was allowed to cool and the organics were removed in vacuo .
  • the aqueous portion was diluted with an equal volume of brine and extracted several times with EtOAc.
  • the reaction mixture was stirred for 16 h at rt, cooled in an ice bath, quenched with 100 mL MeOH, and reduced in vacuo to a smaller volume.
  • the brown suspension was cooled in an ice bath and stirred while 2 L of 5 N HCl was added at a rate to keep the temperature below 25 °C.
  • the suspension was stirred 18 h at rt, then extracted several times with CH2CI2 •
  • the CH2CI2 extracts were combined, washed with brine, dried (Na2S0 ) , and concentrated to give an oily brown solid.
  • the AICI3 (1.2 g, 9 mmol) was stirred in 30 mL of DCE at 0 °C, ethanethiol (1 mL, 14 mmol) was added, and the mixture stirred for 15 min.
  • the 2-[4-[2-(l- piperidinyl)ethoxy]phenyl] -3-(4-methoxyphenyl) -6- methylpyridine (0.5 g, 1.2 mmol) in 20 mL DCE was added dropwise to the reaction mixture and stirred for 2 h as it was allowed to come to rt, then quenched with 25 mL THF at 0 °C, followed by 25 mL 1 N HCl and worked up.
  • the AICI3 (5 g, 38 mmol) was stirred in 100 mL of DCE at 0 °C, ethanethiol (4 mL, 54 mmol) was added, and the mixture stirred for 15 min.
  • the 2-[4-[2-(4- morpholinyl)ethoxy]phenyl] -3-(4-methoxyphenyl) -6- methylpyridine (2.2 g, 5.4 mmol) in 50 mL DCE was added dropwise to the reaction mixture and stirred for 2 h as it was allowed to come to rt, then quenched with 50 mL THF at 0 °C, followed by 50 mL 1 N HCl and worked up.
  • the AICI3 (1.9 g, 14 mmol) was stirred in 50 mL of DCE at 0 °C, ethanethiol (1.5 mL, 20 mmol) was added, and the mixture stirred for 15 min.
  • the 2-[4-[2-(l- piperidinyl)ethoxy]phenyl]-3- (4-methoxyphenyl) -6-(4- fluorophenyl)pyridine (1 g, 2 mmol) in 25 mL DCE was added dropwise to the reaction mixture and stirred for 2 h as it was allowed to come to rt, then quenched with 25 mL THF at 0 °C, followed by 25 mL 1 N HCl and worked up.
  • mice Seventy-five day old female Sprague Dawley rats (weight range of 200 to 225g) were obtained from Charles River Laboratories (Portage, MI) .
  • the animals were either bilaterally ovariectomized (OVX) or exposed to a sham surgical procedure at Charles River Laboratories, and then shipped after one week. Upon arrival, they were housed in metal hanging cages in groups of 3 or 4 per cage and had ad libitum access to food (calcium content approximately 0.5%) and water for one week.
  • Room temperature was maintained at 22.2° ⁇ 1.7° C with a minimum relative humidity of 40%. The photoperiod in the room was 12 hours light and 12 hours dark.
  • EPO Eosinophil Peroxidase Assay.
  • Uteri were kept at 4° C until time of enzymatic analysis.
  • the uteri were then homogenized in 50 volumes of 50 mM Tris buffer (pH - 8.0) containing 0.005% Triton X-100.
  • Tris buffer pH - 8.0
  • Upon addition of 0.01% hydrogen peroxide and 10 mM O-phenylenediamine (final concentrations) in Tris buffer increase in absorbance was monitored for one minute at 450 nm.
  • the presence of eosinophils in the uterus is an indication of estrogenic activity of a compound.
  • the maximal velocity of a 15 second interval was determined over the initial, linear portion of the reaction curve.
  • Table 1 shows comparative results among ovariectomized rats, rats treated with 17 ⁇ - ethynyl estradiol (EE 2 ; an orally available form of estrogen), and rats treated with certain compounds of the present invention.
  • EE 2 caused a decrease in serum cholesterol when orally administered at 0.1 mg/kg/day, it also exerted a stimulatory action on the uterus so that EE 2 uterine weight was substantially greater than the uterine weight of ovariectomized control animals. This uterine response to estrogen is well recognized in the art.
  • the compounds of the present invention generally reduce serum cholesterol compared to the ovariectomized control animals, but uterine weight was only minimally increased to slightly decreased with the majority of the formula compounds tested. Compared to estrogenic compounds known in the art, the benefit of serum cholesterol reduction without adversely affecting uterine weight is quite rare and desirable.
  • estrogenicity also was assessed by evaluating the response of eosinophil infiltration into the uterus. Most of the compounds of the present invention did not cause any increase in the number of eosinophils observed in the stromal layer of ovariectomized rats, while others caused only modest increases. Estradiol caused a substantial, expected increase in eosinophil infiltration.
  • MCF-7 Proliferation Assay MCF-7 breast adenocarcinoma cells (ATCC HTB 22) were maintained in MEM (minimal essential medium, phenol red- free, Sigma, St. Louis, MO) supplemented with 10% fetal bovine serum (FBS) (V/V), L-glutamine (2 mM) , sodium pyruvate (I mM), HEPES ⁇ (N-[2-hydroxyethyl]piperazine-N'-[2- ethanesulfonic acid]10 mM ⁇ , non-essential amino acids and bovine insulin (1 ug/mL) (maintenance medium) .
  • FBS fetal bovine serum
  • L-glutamine 2 mM
  • I pyruvate I pyruvate
  • HEPES ⁇ N-[2-hydroxyethyl]piperazine-N'-[2- ethanesulfonic acid]10 mM ⁇ , non-essential amino acids and bovine insulin (1 u
  • MCF-7 cells were switched to maintenance medium supplemented with 10% dextran coated charcoal stripped fetal bovine serum (DCC-FBS) assay medium) in place of 10% FBS to deplete internal stores of steroids.
  • DCC-FBS dextran coated charcoal stripped fetal bovine serum
  • MCF-7 cells were removed from maintenance flasks using cell dissociation medium (Ca++/Mg++ free HBSS (phenol red-free) supplemented with 10 mM HEPES and 2 mM EDTA) . Cells were washed twice with assay medium and adjusted to 80,000 cells/mL.
  • Test 1 Between 3 and 20 women having uterine fibrosis are administered a compound of the present invention.
  • the amount of compound administered is from 0.1 to 1000 mg/day, and the period of administration is 3 months.
  • the women are observed during the period of administration, and up to 3 months after discontinuance of administration, for effects on uterine fibrosis.
  • animals in Groups 1 and 2 receive intraperitoneal injections of water for 14 days whereas animals in Group 3 receive intraperitoneal injections of 1.0 mg of a compound of the present invention per kilogram of body weight for the same duration.
  • each female is sacrificed and the endometrial explants, adrenals, remaining uterus, and ovaries, where applicable, are removed and prepared for histological examination. The ovaries and adrenals are weighed.
  • Autographs of endometrial tissue are used to induce endometriosis in rats and/or rabbits.
  • Female animals at reproductive maturity undergo bilateral oophorectomy, and estrogen is supplied exogenously thus providing a specific and constant level of hormone.
  • Autologous endometrial tissue is implanted in the peritoneum of 5-150 animals and estrogen supplied to induce growth of the explanted tissue.
  • Treatment consisting of a compound of the present invention is supplied by gastric gavage on a daily basis for 3-16 weeks, and implants are removed and measured for growth or regression. At the time of sacrifice, the intact horn of the uterus is harvested to assess status of endometrium.
  • Tissue from human endometrial lesions is implanted into the peritoneum of sexually mature, castrated, female, nude mice. Exogenous estrogen is supplied to induce growth of the explanted tissue.
  • the harvested endometrial cells are cultured in vitro prior to implantation. Treatment consisting of a compound of the present invention supplied by gastric gavage on a daily basis for 3-16 weeks, and implants are removed and measured for growth or regression. At the time of sacrifice, the uteri are harvested to assess the status of the intact endometrium.
  • Tissue from human endometrial lesions is harvested and maintained in vi tro as primary nontransformed cultures. Surgical specimens are pushed through a sterile mesh or sieve, or alternately teased apart from surrounding tissue to produce a single cell suspension. Cells are maintained in media containing 10% serum and antibiotic. Rates of growth in the presence and absence of estrogen are determined. Cells are assayed for their ability to produce complement component C3 and their response to growth factors and growth hormone. In vi tro cultures are assessed for their proliferative response following treatment with progestins, GnRH, a compound of the invention, and vehicle. Levels of steroid hormone receptors are assessed weekly to determine whether important cell characteristics are maintained in vi tro . Tissue from 5-25 patients is utilized.
  • Compounds of the present invention have capacity to inhibit aortal smooth cell proliferation. This can be demonstrated by using cultured smooth cells derived from rabbit aorta, proliferation being determined by the measurement of DNA synthesis. Cells are obtained by explant method as described in Ross, J. of Cell Bio. 50: 172 (1971) . Cells are plated in 96 well microtiter plates for five days. The cultures become confluent and growth arrested.
  • the cells are then transferred to Dulbecco's Modified Eagle's Medium (DMEM) containing 0.5 - 2% platelet poor plasma, 2 mM L- glutamine, 100 U/ml penicillin, 100 mg ml streptomycin, 1 mC/ml 3 H-thymidine, 20 ng/ml platelet-derived growth factor, and varying concentrations of the present compounds.
  • Stock solution of the compounds is prepared in dimethyl sulphoxide and then diluted to appropriate concentration_ (0.01 - 30 mM) in the above assay medium.
  • Cells are then incubated at 37° C. for 24 hours under 5% C0 2 /95% air. At the end of 24 hours, the cells are fixed in methanol. 3 H thymidine incorporation in DNA is then determined by scintillation counting as described in Bonin, et al . , Exp. Cell Res. 181: 475-482 (1989).
  • Inhibition of aortal smooth muscle cell proliferation by the compounds of the present invention are further demonstrated by determining their effects on exponentially growing cells.
  • Smooth muscle cells from rabbit aortae are seeded in 12 well tissue culture plates in DMEM containing 10% fetal bovine serum, 2 mM L-glutamine, 100 U/ml penicillin, and 100 mg/ml streptomycin. After 24 hours, the cells are attached and the medium is replaced with DMEM containing 10% serum, 2 mM L-glutamine, 100 U/ml penicillin, 100 mg/ml streptomycin, and desired concentrations of the compounds. Cells are allowed to grow for four days. Cells are treated with trypsin and the number of cells in each culture is determined by counting using a ZM-Coulter counter. Activity in the above tests indicates that the compounds of the present invention are of potential in the treatment of restenosis.
  • estrogen includes steroidal compounds having estrogenic activity such as, for example, 17 ⁇ -estradiol, estrone, conjugated estrogen (Premarin®) , equine estrogen, 17 ⁇ -ethynyl estradiol, and the like.
  • progestin includes compounds having progestational activity such as, for example, progesterone, norethynodrel, norgestrel, megestrol acetate, norethindrone, and the like.
  • Estrogen-based agents include, for example, ethynyl estrogen (0.01 - 0.03 mg/day), mestranol (0.05 - 0.15 mg/day), and conjugated estrogenic hormones such as Premarin® (Wyeth-Ayerst; 0.3 - 2.5 mg/day).
  • Progestin- based agents include, for example, medroxyprogesterone such as Provera® (Upjohn; 2.5 -10 mg/day), norethynodrel (1.0 - 10.0 mg/day), and norethindrone (0.5 - 2.0 mg/day).
  • a typical daily dose will contain a nontoxic dosage level of from about 5 mg to about 1000 mg/day of a compound of the present invention.
  • Preferred daily doses generally will be from about 15 mg to about 500 mg/day.
  • the compounds also can be formulated as elixirs or solutions for convenient oral administration or as solutions appropriate for parenteral administration, for example, by intramuscular, subcutaneous or intravenous routes. Additionally, the compounds are well suited to formulation as sustained release dosage forms and the like.
  • the formulations can be so constituted that they release the active ingredient only or preferably in a particular physiological location, possibly over a period of time.
  • the coatings, envelopes, and protective matrices may be made, for example, from polymeric substances or waxes.
  • Hard gelatin capsules are prepared using the following:
  • the medicament is passed through a No. 45 mesh U.S. sieve and mixed with the sodium carboxymethyl cellulose and syrup to form a smooth paste.
  • the benzoic acid solution, flavor, and color are diluted with some of the water and added, with stirring. Sufficient water is then added to produce the required volume.
  • An aerosol solution is prepared containing the following ingredients:
  • Suppositories are prepared as follows:
  • the active ingredient is passed through a No. 60 mesh U.S. sieve and suspended in the saturated fatty acid glycerides previously melted using the minimal necessary heat. The mixture is then poured into a suppository mold of nominal 2 g capacity and allowed to cool.
  • An intravenous formulation is prepared as follows:
  • Active ingredient 50 Premarin 1 Avicel pH 101 50 Starch 1500 117.50 Silicon Oil 2 Tween 80 0.50 Cab-O-Sil 0.25
  • Active ingredient 50 Norethynodrel 5 Avicel pH 101 82.50 Starch 1500 90 Silicon Oil 2 Tween 80 0.50
  • Active ingredient 50 Premarin 1 Corn Starch NF 50 Povidone, K29-32 6 Avicel pH 101 41.50 Avicel pH 102 136.50 Crospovidone XL10 2.50 Magnesium Stearate 0.50 Cab-O-Sil 0.50

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Medicinal Chemistry (AREA)
  • Organic Chemistry (AREA)
  • Epidemiology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Diabetes (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Engineering & Computer Science (AREA)
  • Obesity (AREA)
  • Hematology (AREA)
  • Endocrinology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Pyridine Compounds (AREA)

Abstract

Cette invention concerne des composés de Formule (I), ou n est 2 ou 3; R désigne un groupe diméthylamino, diéthylamino, pipéridinyle-1, pyrrolidinyle-1, morpholinyle-4, ou hexaméthylène-imino-1; R1 désigne l'hydrogène, un groupe alkyle inférieur C¿1?-C4, phényle, ou phényle mono ou bi-substitué, dans lequel chaque substituant est indépendamment l'un de l'autre un halogène, un groupe méthyle, hydroxy, alcoxy C1-C6, benzyloxy, alcanoyloxy C1-C6 ou benzoyloxy, le benzoyloxy substitué portant 1 à 3 substituants dont chacun est indépendamment l'un de l'autre un halogène, un groupe alkyle inférieur C1-C4, alcoxy inférieur C1-C4, alcoxycarbonyloxy C1-C5 ou alkylsulfonyloxy C4-C6; R?2¿ désigne l'hydrogène, un groupe hydroxy, alkoxy C¿1?-C6, benzyloxy, alcanoyloxy C1-C6 ou benzoyloxy, le benzoyloxy portant 1 à 3 substituants dont chacun est indépendamment l'un de l'autre un halogène, un groupe alkyle inférieur C1-C4, alcoxy inférieur C1-C4, alcoxycarbonyloxy C1-C5, ou alkylsulfonyloxy C4-C6. L'invention concerne également des sels d'addition d'acides pharmaceutiquement acceptables destinés au traitement des symptômes post-ménopausiques tels qu'ostéoporose et troubles cardio-vasculaires (hyperlipidémie par exemple), etc.
PCT/US1996/011958 1995-07-27 1996-07-18 Composes pyridiniques, intermediaires, processus industriels, compositions et procedes Ceased WO1997004778A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
JP9507648A JPH11510163A (ja) 1995-07-27 1996-07-18 ピリジン化合物、中間体、製法、組成物及び方法
AU65030/96A AU6503096A (en) 1995-07-27 1996-07-18 Pyridine compounds, intermediates, processes, compositions, and methods

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US158895P 1995-07-27 1995-07-27
US60/001,588 1995-07-27

Publications (1)

Publication Number Publication Date
WO1997004778A1 true WO1997004778A1 (fr) 1997-02-13

Family

ID=21696833

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1996/011958 Ceased WO1997004778A1 (fr) 1995-07-27 1996-07-18 Composes pyridiniques, intermediaires, processus industriels, compositions et procedes

Country Status (4)

Country Link
JP (1) JPH11510163A (fr)
AU (1) AU6503096A (fr)
CA (1) CA2227877A1 (fr)
WO (1) WO1997004778A1 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6174901B1 (en) * 1998-12-18 2001-01-16 Amgen Inc. Substituted pyridine and pyridazine compounds and methods of use
US6956132B2 (en) 2001-08-03 2005-10-18 Ishihara Sangyo Kaisha. Ltd. Process for producing 2-phenylacetophenone derivatives and precursors therefor
US7271266B2 (en) 2002-03-28 2007-09-18 Merck & Co., Inc. Substituted 2,3-diphenyl pyridines

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3535330A (en) * 1968-04-29 1970-10-20 Sandoz Ag 2,6-diphenyl - 4 - (p-(dilower-alkyl amino lower - alkoxy)phenyl)pyridines and derivatives thereof
US3535329A (en) * 1969-03-07 1970-10-20 Sandoz Ag Certain 2-(4-(dilower alkylaminoloweralkoxy)phenyl)-4,6-diphenyl pyridines and derivatives thereof

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3535330A (en) * 1968-04-29 1970-10-20 Sandoz Ag 2,6-diphenyl - 4 - (p-(dilower-alkyl amino lower - alkoxy)phenyl)pyridines and derivatives thereof
US3535329A (en) * 1969-03-07 1970-10-20 Sandoz Ag Certain 2-(4-(dilower alkylaminoloweralkoxy)phenyl)-4,6-diphenyl pyridines and derivatives thereof

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6174901B1 (en) * 1998-12-18 2001-01-16 Amgen Inc. Substituted pyridine and pyridazine compounds and methods of use
US6956132B2 (en) 2001-08-03 2005-10-18 Ishihara Sangyo Kaisha. Ltd. Process for producing 2-phenylacetophenone derivatives and precursors therefor
US7271266B2 (en) 2002-03-28 2007-09-18 Merck & Co., Inc. Substituted 2,3-diphenyl pyridines

Also Published As

Publication number Publication date
CA2227877A1 (fr) 1997-02-13
JPH11510163A (ja) 1999-09-07
AU6503096A (en) 1997-02-26

Similar Documents

Publication Publication Date Title
US7501441B1 (en) Naphthyl compounds, intermediates, processes, compositions, and methods
US6268361B1 (en) Naphthyl compounds, intermediates, compositions, and methods
US5728724A (en) Benzothiophene compounds
US5643896A (en) 2-Benzyl-3-arylbenzothiophenes
EP0806420B1 (fr) Benzothiophène, formulations les contenant et procédés
US5811421A (en) Naphthyl and dihydronaphthyl intermediates, compounds, compositions, and methods
US5622974A (en) α-substituted-3-benzyl-benzofurans
US5750708A (en) Pyridine compounds intermediates processes compositions and methods
US6451817B1 (en) Alpha-substituted-1-benzyl-napthyls
US5998443A (en) Phosphorous containing benzothiophenes for treating estrogen deficiency
US5705507A (en) Aplha-substituted-3-benzyl-benzofurans
US6417199B1 (en) 3-benzyl-benzothiophenes
WO1997004778A1 (fr) Composes pyridiniques, intermediaires, processus industriels, compositions et procedes
GB2303628A (en) 5-Hydroxy- and 5-phenyl-substituted 1-[4-(aminoalkoxy)phenyl]-2-phenylbenzene derivatives for the treatment of post menopausal symptoms and intermediates
US6384053B1 (en) α-substituted-1-benzyl-napthyls
US6653328B1 (en) 3-benzyl-benzothiophenes
IL128310A (en) Intermediates for naphthyl compounds and processes for their preparation

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AL AM AU AZ BB BG BR BY CA CN CU CZ EE GE HU IL IS JP KE KG KP KR KZ LK LR LS LT LV MD MG MK MN MW MX NO NZ PL RO RU SD SG SI SK TJ TM TR TT UA UG US UZ VN AM AZ BY KG KZ MD RU TJ TM

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): KE LS MW SD SZ UG BF BJ CF CG CI CM GA GN ML MR NE SN TD TG

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
ENP Entry into the national phase

Ref document number: 2227877

Country of ref document: CA

Ref country code: CA

Ref document number: 2227877

Kind code of ref document: A

Format of ref document f/p: F

ENP Entry into the national phase

Ref country code: JP

Ref document number: 1997 507648

Kind code of ref document: A

Format of ref document f/p: F