CN1209746A - Method of inhibiting musculoaponeurotic fibromatoses (desmoid tumors) - Google Patents

Method of inhibiting musculoaponeurotic fibromatoses (desmoid tumors) Download PDF

Info

Publication number
CN1209746A
CN1209746A CN97191916A CN97191916A CN1209746A CN 1209746 A CN1209746 A CN 1209746A CN 97191916 A CN97191916 A CN 97191916A CN 97191916 A CN97191916 A CN 97191916A CN 1209746 A CN1209746 A CN 1209746A
Authority
CN
China
Prior art keywords
chemical compound
preparation
cell
acid
milligram
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.)
Pending
Application number
CN97191916A
Other languages
Chinese (zh)
Inventor
M·L·布兰迪
F·托内利
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
Priority claimed from GBGB9603148.9A external-priority patent/GB9603148D0/en
Application filed by Eli Lilly and Co filed Critical Eli Lilly and Co
Publication of CN1209746A publication Critical patent/CN1209746A/en
Pending legal-status Critical Current

Links

Classifications

    • 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
    • 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
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
    • A61K31/4535—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a heterocyclic ring having sulfur as a ring hetero atom, e.g. pizotifen
    • 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
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • 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
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A61K31/4025—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil not condensed and containing further heterocyclic rings, e.g. cromakalim
    • 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
    • A61K31/55—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Epidemiology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Immunology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Hydrogenated Pyridines (AREA)
  • Pyrrole Compounds (AREA)

Abstract

A method of inhibiting musculoaponeurotic fibromatoses comprising administering to a mammal in need thereof an effective amount of a compound having formula (I), wherein R<1> and R<3> are independently hydrogen, -CH3, (a) ou (b), wherein Ar is optionally substituted phenyl; R<2> is selected from the group consisting of pyrrolidine, hexamethyleneimino, and piperidino; or a pharmaceutically acceptable salt or solvate thereof.

Description

The method that suppresses aponeurosis fibromatosis (fibroma durum)
Background of invention
The aponeurosis fibromatosis is the local invasive dysplasia of the connective tissue damage of a class non-metastatic.Such disease comprises nodular fasciitis, sole of the foot fibromatosis and before had been divided into the damage of fibroma durum.
Great majority " hard fibre " damage relates to skeletal muscle and relevant fascia layer.Their the most normal stomach walls that betides phenolics or pregnant back women; They also are common in the male and betide the outer position of abdominal part, comprise head and cervical region, thigh and shoulder.
Damage also accidentally in operative scar and mesentery, a kind of common form is relevant with the Gardner syndrome.The Therapeutic Method of suggestion is to excise along normal structure edge large tracts of land.But brothers and main blood vessel and nerve should be without prejudice, even might recur.Common local recurrence also often needs excision again.X-ray therapy is also effective to these damages.Some situation adopts the tamoxifen treatment effectively.In any case but, still need other therapy.
Summary of the invention
The invention provides the method that suppresses mammal aponeurosis fibromatosis, this method is passed through the formula I compound or pharmaceutically acceptable salt thereof of the administration effective dose of this inhibition of need or solvate realization,
R wherein 1And R 3Be independently hydrogen ,-CH 3,
Figure A9719191600032
Alkyl) or
Figure A9719191600033
Wherein Ar is the optional phenyl that replaces;
R 2Be selected from pyrrolidino, hexamethyleneimino and piperidino.
Detailed Description Of The Invention
The present invention relates to the discovery of one group of 2-phenyl-3-aroyl benzothiophene (benzothiophene) chemical compound, formula I chemical compound wherein can be used for suppressing the aponeurosis fibromatosis.
Using method provided by the invention is by giving the people need this inhibition and use the formula I compound or pharmaceutically acceptable salt thereof or solvate being achieved, and this chemical compound is effective to suppressing the aponeurosis fibromatosis.Term " inhibition " comprises its common art-recognized meanings, comprises the symptom or the effect of prevention, prevention, restriction and alleviation, prevention or reverse disease progression, the order of severity or generation.
Raloxifene, a kind of wherein R 1And R 3Be hydrogen and R 2Being the hydrochlorate of the formula I chemical compound of the present invention of piperidino, is that a kind of nuclear is regulated molecule.Raloxifene has shown the molecule that combines and be considered at first have estrogen antagonist function and pharmacotoxicological effect with estrogen receptor, because its blocking-up estrogen activates the ability of uterine cancer cell and estrogen-dependent breast carcinoma.Really, raloxifene is blocked estrogenic effect in some cell; But in the cell of other type, raloxifene activates identical gene and demonstrates identical pharmacological properties with estrogen, as causes osteoporosis, hyperlipidemia.As a result, raloxifene is considered to have a kind of estrogen antagonist of mixing excitement-antagonist properties.Think that now the peculiar property different with estrogen that raloxifene shows is owing to activation and/or the inhibitory action of raloxifene-estrogen receptor complex to the uniqueness of range gene function, different with estrogen-estrogen receptor complex to the activation and/or the inhibitory action of gene.Therefore, though raloxifene and estrogen utilization and competing phase receptor together, and be not easy to predict the two pharmacology result to Gene regulation, they respectively have characteristics.
In general, chemical compound and usual excipients, diluent or carrier preparation are pressed into tablet; Or be formulated as convenient oral elixir or solution administration or through intramuscular or intravenous route administration.But these chemical compound percutaneous dosings, and can be mixed with slow release formulation etc.
The chemical compound that uses in the method for the present invention can prepare in accordance with known methods, and as those methods that describes in detail in United States Patent (USP) 4133814,4418068 and 4380635, these documents are incorporated herein by reference.Usually, from being with benzo [b] thiophene of 6-hydroxyl and 2-(4-hydroxy phenyl).With starting compound protected, acidylate, deprotection forms the formula I chemical compound then.Provided the preparation embodiment of these chemical compounds in the above-mentioned United States Patent (USP).Arbitrarily the phenyl that replaces comprises phenyl and by one or two C 1-C 6Alkyl, C 1-C 4Alkoxyl, hydroxyl, nitro, chlorine, the methyl substituted phenyl of fluorine or three (chlorine or fluorine).
Chemical compound and various organic and inorganic bronsted lowry acids and bases bronsted lowry that the inventive method is used form pharmaceutically useful bronsted lowry acids and bases bronsted lowry addition salts, comprise physiologically acceptable salt commonly used in the pharmaceutical chemistry.This class salt also is a part of the present invention.The typical inorganic acid that is used to form this class salt comprises hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, sulphuric acid, phosphoric acid and hypophosphoric acid etc.Also can use alkanoic acid, hydroxyl alkane acid and the hydroxyl chain docosandioic acid, the aromatic carboxylic acid that replace by for example aliphatic list of organic acid or dicarboxylic acids, phenyl, the derive salt of formation such as aliphatic series and aromatic sulfonic acid.This class officinal salt comprises acetate, phenylacetate, trifluoroacetate, acrylates, Ascorbate, benzoate, chloro benzoate, dinitro-benzoate, hydroxy benzoate, methoxybenzoic acid salt, ar-Toluic acid salt, acetoxybenzoic acid salt, naphthalene-2-benzoate, bromide, isobutyrate, phenylbutyric acid salt, beta-hydroxy-butanoic acid salt, butine-1, the 4-diacid salt, hexin-1, the 4-diacid salt, caprate, caprylate, chloride, cinnamate, citrate, formates, fumarate, glycollate, enanthate, hippurate, lactate, malate, maleate, hydroxymaleic acid salt, malonate, mandelate, mesylate, nicotinate, .gamma.-pyridinecarboxylic acid salt, nitrate, oxalates, phthalate, terephthalate, phosphate, dibasic alkaliine, dihydric phosphate, metaphosphate, pyrophosphate, propiolate, propionate, phenylpropionic acid salt, Salicylate, sebacate, succinate, suberate, sulfate, disulfate, pyrosulfate, sulphite, dithionite, sulfonate, benzene sulfonate, brosylate, closilate, esilate, 2 monohydroxy esilates, mesylate, naphthalene-1-sulfonate, naphthalene-2-sulfonic acid salt, tosilate, xylenesulfonate, tartrate etc.Preferred salt is hydrochlorate.
The pharmaceutically acceptable acid addition salts is normally by formula I chemical compound and equimolar amounts or excessive acid reaction formation.General with the cosolvent of reactant, as mixing in ether or the benzene at them.Usually be precipitated out from solution at one hour or 10 days inner salts, this salt can be after filtration or is removed to desolvate with conventional method and be separated.
The alkali that is used to form salt comprises ammonium hydroxide and alkali metal and alkaline earth metal hydroxide, carbonate and aliphatic series and primary, secondary, tertiary amine, aliphatic diamine.The alkali that is particularly suitable for preparing addition salts comprises ammonium hydroxide, potassium carbonate, methylamine, diethylamine, ethylenediamine and cyclohexylamine.
Compare with their chemical compound of deriving, the common dissolubility of officinal salt improves, and therefore, they usually are more suitable for obtaining liq or Emulsion.
Pharmaceutical preparation can be adopted the methods known in the art preparation.For example, chemical compound and usual excipients, diluent or carrier preparation can be formed tablet, capsule, suspension, powder etc.The example that is applicable to excipient, diluent and the carrier of this class preparation comprises following: filler and extender, as starch, saccharide, mannitol and siliceous derivant; Binding agent is as carboxymethyl cellulose and other cellulose derivative, alginate, gelatin and polyvinylpyrrolidone; Wetting agent is as glycerol; Disintegrating agent is as calcium carbonate and sodium bicarbonate; Postpone dissolved reagent, as paraffin; Absorption enhancer is as quaternary ammonium compound; Surfactant, as spermol, glyceryl monostearate; Absorption carrier is as Kaolin and bentonite; And lubricant, as Talcum, calcium stearate and magnesium stearate and solid polyethylene glycol.
Also chemical compound can be mixed with elixir or the solution that makes things convenient for oral administration, perhaps be mixed with and be suitable for parenteral route, as the solution of intramuscular, subcutaneous or intravenous route administration.In addition, these chemical compounds are suitable for being mixed with slow release formulation etc. very much.Only preparation can be mixed with or preferably at intestinal specific part release of active ingredients in a period of time.Also can have coating, seal and protectiveness substrate, as using polymer or wax.
According to the present invention, the given dose that suppresses the required formula I chemical compound of aponeurosis fibromatosis or other purposes disclosed herein will depend on the order of severity of disease, route of administration and the other factors that is determined by the clinicist.Usually, acceptable effective every day, dosage was about 1000 mg/day of about 0.1-, and more typical is about 200 mg/day of about 50-.Be effectively to suppress the aponeurosis tumor, to this inhibition of need by treatment target, but this dosage be administered once to about three times every day, or as required administration more times.
The formula I chemical compound is usually preferably with the acid-addition salts form administration, as having basic group as the practice commonly used of the medicine of piperidino ring.By this compounds of oral administration also is useful.For this reason, can utilize following peroral dosage form.
For topical, can be according to the technology preparation chemical compound that directly applies to the surface known in the art.Conventionally form for this purpose comprises ointment, lotion, paste, gel, spray and aerosol.The weight percentage of The compounds of this invention in topical formulations depends on various factors, but is generally the 0.5%-95% of total formulation weight amount, is typically 1-25% (weight).
Compositions can be moisture or anhydrous solution or dispersion, perhaps is emulsion or form of suspension.
These compositionss can contain pharmaceutically suitable carrier commonly known in the art and adjuvant.For example, be preparation solution, can use the acceptable organic solvent of one or more physiology, outside dewatering, also be selected from the C of acetone, ethanol, isopropyl alcohol, glycol ether (commercially available prod that is called " Dowanol " as name), polyethylene glycols, polyoxyethylene, short chain acids 1-C 4Arrcostab (preferred ethyl lactate or isopropyl lactate), fatty acid triglycercide (commercially available prod that is called " Miglyol " as name), isopropyl myristate and animal, mineral and vegetable oil and polysiloxanes.
Compositions of the present invention also can contain thickening agent, as cellulose and/or cellulose derivative.They also can contain natural gum, as Tragacanth, guar gum or carob or arabic gum, and perhaps Polyethylene Glycol, bentonite and montorillonite clay etc.
The lid human relations form that mainly is suitable for topical application is cream, milk, gel, dispersion liquid or microemulsion form, be more or less thickening lotion, impregnated pads, ointment or bar-shaped dosage form, perhaps be the spraying or the aerosol or the soap blank form of form of foam.
Preparation
In the infra series preparation, " active component " is meant the formula I chemical compound.
Preparation 1: gelatine capsule
Adopt following ingredients to prepare hard gelatin capsule:
Composition Amount (milligram/capsule)
Active component starch, NF starch, but the flowing powder silicone liquid, 350 centistokes ??????0.1-1000 ???????0-650 ???????0-650 ????????0-15
Composition is mixed and mistake 45 order U.S. sieves, be filled in the hard gelatin capsule then.The example of the specific raloxifene capsule preparations of preparation comprises following ingredients: preparation 2: raloxifene capsule
Composition Amount (milligram/capsule)
Raloxifene starch, NF starch, but the flowing powder silicone liquid, 350 centistokes ?????????1 ????????112 ???????225.3 ????????1.7
Preparation 3: raloxifene capsule
Composition Amount (milligram/capsule)
Raloxifene starch, NF starch, but the flowing powder silicone liquid, 350 centistokes ?????????5 ????????108 ???????225.3 ????????1.7
Preparation 4: raloxifene capsule
Composition Amount (milligram/capsule)
Raloxifene starch, NF starch, but the flowing powder silicone liquid, 350 centistokes ?????????10 ????????103 ???????225.3 ????????1.7
Preparation 5: raloxifene capsule
Composition Amount (milligram/capsule)
Raloxifene starch, NF starch, but the flowing powder silicone liquid, 350 centistokes ?????????50 ????????150 ????????397 ????????3.0
Above-mentioned concrete prescription can change according to the reasonable change of defined.Adopt following compositions to prepare tablet: preparation 6: tablet
Composition Amount (milligram/sheet)
Active component microcrystalline Cellulose silicon dioxide, vaporific stearic acid ?????0.1-1000 ??????0-650 ??????0-650 ???????0-15
Each composition mixing and compacting is in blocks.Perhaps, be prepared as follows the tablet that respectively contains 0.1-1000 milligram active component: system 7: tablet
Composition Amount (milligram/sheet)
Active component starch microcrystalline cellulose polyvinylpyrrolidone (10% aqueous solution) sodium carboxymethylcellulose dolomol talcum ?????0.1-1000 ????????45 ????????33 ????????4 ???????4.5 ???????0.5 ????????1
Active component, starch and cellulose are crossed 45 order U.S. sieves and fully mixing.With polyvinylpyrrolidonesolution solution and gained powder mixes, cross 14 order U.S. sieves then.The granule that so obtains in 50-60 ℃ of drying, is crossed 18 order U.S. sieves then.Sodium carboxymethyl cellulose, magnesium stearate and Talcum by 60 order U.S. sieves in advance is added in this granule, and after the mixing, tabletting obtains tablet on tablet machine.
Be prepared as follows per 5 milliliters of suspensions that contain 0.1-1000 milligram medicine:
Preparation 8: suspension
Composition Amount (milligram/5 milliliters)
Active fully sodium carboxymethyl cellulose syrup benzoic acid solution aromatic coloring agent pure water adds to 0.1-1000 50 milligrams 1.25 milligrams 0.10 milligram an amount of 5 milliliters of milligrams
Medicine is mixed the soft and smooth pastel of formation by 45 order U.S. sieves and with sodium carboxymethyl cellulose and syrup.Benzoic acid solution, aromatic and coloring agent are also under agitation added with a certain amount of water dilution.Add enough water then to volume required.Prepare following topical composition: preparation 9
Composition Amount (milligram/5 milliliters)
Hydroxypropyl cellulose active component isopropyl alcohol, an amount of 1.5 gram 1.5-30 gram 100 grams
Preparation 10
Composition Amount (milligram/5 milliliters)
Hydroxypropyl cellulose ethyl lactate active component isopropyl alcohol, an amount of 1.5 restrain 15.0 gram 1.5-30 grams, 100 grams
Preparation 11
Composition Amount (milligram/5 milliliters)
Hydroxypropyl cellulose Yoshinox BHT active component isopropyl alcohol, an amount of 1.0 restrain 0.02 gram 1.5-25 gram, 100 grams
Preparation 12
Composition Amount (milligram/5 milliliters)
Hydroxypropyl cellulose Yoshinox BHT C 8-C 12Fatty acid triglycercide active component isopropyl alcohol, an amount of 1.5 restrain 0.01 gram, 10.0 gram 1.5-30 grams, 100 grams
Preparation 9-12 is a gel form.Preparation 13
Composition Amount (milligram/5 milliliters)
Isopropyl alcohol active component C 8-C 12Fatty acid triglycercide 46.0 gram 1.0-15 gram 49.0 grams
Preparation 14
Composition Amount (milligram/5 milliliters)
Ethanol ethyl lactate active component C 8-C 12Fatty acid triglycercide 69.0 restrain 10.0 gram 1.5-20 grams, 30.0 grams
Preparation 15
Composition Amount (milligram/5 milliliters)
Isopropyl alcohol acetone ethyl lactate active component C 8-C 12Fatty acid triglycercide 47.0 restrain 10.0 grams, 10.0 gram 1-15 grams, 30.0 grams
Preparation 16
Composition Amount (milligram/5 milliliters)
The butylated hydroxy-methylbenzene active component of ethanol 95.08 restrain 0.02 gram 1.5-25 gram
Preparation 13,14,15 and 16 is lotion form.Preparation 17
Composition Amount (milligram/5 milliliters)
White vaseline liquid paraffin fully refined paraffin wax active component 50.0 restrain 15.0 grams, 32.0 gram 1-20 grams
Preparation 18
Composition Amount (milligram/5 milliliters)
White vaseline liquid paraffin fully refined paraffin wax active component 50.0 restrain 13.0 grams, 32.0 gram 1-20 grams
Preparation 17 and 18 is bar-shaped dosage form.
Fibroma durum is rare fibre source non-metastatic tumor.Clinical verification shows that steroid hormone may work in the natural history of this class tumor: it is mainly seen among the women of child-bearing age patient, and the degeneration of this class tumor is relevant with the estrogen antagonist therapy with menopause.
The purpose of this research is the estrogen receptor of confirming in the fibroma durum primary cell, estimates the effect of formula I chemical compound to fibroma durum cell in the primary culture.
Because fibroma durum betides among familial adenomatous polyposis (FAP) patient sometimes, it can be degenerated in colon or rectal cancer patient, and we have tested the fibroblastic cell growth inhibition of formula I a chemical compound to gland cell system (HCT8) and colon cancer patient biopsy samples.
Chemical compounds I a is R wherein 2Be pyrrolidino and R 1And R 3It is the formula I chemical compound of hydrogen.
Use intact cell to carry out associativity research.The fibroma durum cell is tiled on 6 well culture plates of growth medium (being supplemented with the Ham F12 of the Coon modification of 10%FCS).After 24 hours, do not replace this growth medium, cell was kept 24 hours at starvation with not adding phenol red stable state culture medium.Then with cell with 1 milliliter do not add phenol red but contain the culture medium (binding buffer liquid) of 25mM HEPES and 0.5%EtOH and increased concentrations (0.05-10nM) [ 3H] 17 β E 2(wherein contain or do not contain 500 times of excessive unlabelled 17 β E 2With formula I a chemical compound) cultivated 1 hour.After the cultivation, cell is with 800 microlitre binding buffer liquid washed twice and use 1N NaOH in 70 ℃ of cracking 30 minutes.Then 4N HCl is added in every hole so that neutralization.Measure radioactivity with the liquid scintillation spectrometry method.Estimate ER binding affinity and binding capacity by the Scatchard analytic process.
Institute subsequently all carries out under 0-4 ℃ in steps.Be organized in the buffer in polytron homogenizer homogenize twice with what grind, each 10 seconds, there is 30 seconds cool time the centre, and the composition of buffer is: 10mM Tris-HCl, 5mMEDTA, 10mM sodium molybdate, 10mM 1,4-Dithioerythritol, 10% glycerol (v/v), pH 7.4.With homogenate centrifugal 20 minutes at 7000g, abandon precipitate, supernatant centrifugal again 60 minutes in 105000g obtains being used for the cytosol that estrogen receptor analysis is used.Cytosol is diluted to 1-2 milligram albumen/milliliter.Measure cytosol albumen according to the Bradford method.For carrying out estrogen receptor analysis, with cytosol in 4 ℃ and concentration range be 0.05-5nM [ 3H] 17 β E 2And with or not with 500 times of excessive unlabelled 17 β E 2Cultivate together with formula I a chemical compound.Estimate ER binding affinity and binding capacity by the Scatchard analytic process.
With cell with every hole 8 * 10 4The density of individual cell is tiled in 6 well culture plates of growth medium.After 24 hours, with not adding phenol red but be supplemented with 0.1%DMF, 0.1%EtOH and variable concentrations formula I a chemical compound (2 * 10 -5M, 10 -5M, 5 * 10 -6M, 10 -6M) growth medium stimulates.
With cell culture 6 days, move it with trypsin/edta solution, estimate growing state by microscopic counting then.Use same procedure to estimate colon cancer initial stage fibroblast and HCT8 cell line: this cell line is cultivated in the RPM I and was cultivated 4 days after stimulation.
With I Collagen Type VI and the cellular layer in enzyme-linked immunosorbent assay (ELISA) the mensuration culture medium.In brief, cell was cultivated 24 hours in containing the Coon modification Ham F12 culture medium that does not add additional nutrient of 50 mcg/ml ascorbic acid and 100 mcg/ml β-An Jibingjing base (propionitryl) fumarates.
The results culture medium is also used suitably dilution of 0.1M carbonate buffer (pH9.6), wrap by the ELISA culture plate with it then, spend the night in 4 ℃, for making nonspecific binding site saturated, the ELISA culture plate was cultivated 1.5 hours in 37 ℃ in the PBS that contains 5% milk powder (PBS Blotto), cultivated 2 hours in 37 ℃ with the PBS Blotto that contains specific polyclonal antibody, containing goat anti-rabbit igg-alkaline phosphatase conjugation complex (Sigma Chemical Co., St.Louis cultivated 1.5 hours in 37 ℃ among PBS Blotto MO).
Use 50 mcg/ml Mg++ and 1 mg/ml p-nitrophenyl phosphate ester substrate then, sample is contacted in room temperature with 10% diethanolamine (pH9.8) as alkali phosphatase.Read optical density in 405 nanometers, and according to the standard curve calculating concentration.The I Collagen Type VI of results cell monolayer and ultrasonic mensuration cell in the 0.5N sodium hydroxide.Then in 0.1M carbonate buffer (pH9.6) the diluting cells extract and with its bag by the ELISA culture plate.With standard substance and sample replicate analysis three times.The result represents with microgram albumen/microgram cell DNA.Dna content is measured through spectrofluorimetry.
Former generation the fibroma durum cell and refrigerated fibroma durum sample in [ 3H] 17 β E 2Carry out the associativity test as part.
In two kinds of tests, [ 3H] 17 β E 2Replaced by excessive two kinds of unlabelled estrogen and chemical compounds I a by 500 in conjunction with only a small amount of (about 10%).Use a computer in conjunction with program LIGAND to [ 3H] 17 β E 2The Scatchard that binding data carries out analyzes (Munson P.J., Rodbard D.Anal.Biochem.1980; 107:220-39) show in the two kind different cytosols of three kinds of different cultures neutralizations and all have ER by the biopsy samples preparation of fibroma durum.
In growth analysis, when fibroma durum primary cell when the formula I a of various concentration chemical compound contacts is upset.To be cell growth be suppressed (table 1) with the increase of chemical compounds I a concentration to the result.Use HCT8 cell line (table 2) and colon cancer fibroblast (table 3) also to obtain similar results.
It is 10 by concentration that the fibroma durum cell relies on mode with dosage -5M, 5 * 10 -6M, 10 -6The formula I a chemical compound of M suppresses, in 10 -5Obtain maximum inhibitory action (table 4) during M concentration.
Formula I a chemical compound only under high concentration (500 times excessive) alternative [ 3H] 17 β E 2With the fibroma durum tissue bond.
Formula I a chemical compound can obviously suppress the fibroma durum cell proliferation under millimolar concentration.In addition, under similar concentration, this chemical compound also suppresses the epithelium and the fibroblasts proliferation that are obtained by human colon carcinoma.
The generation of I Collagen Type VI also obviously reduces owing to chemical compounds I a in the fibroma durum cell in primary culture.
Under the transfection experimental condition (electroporation, the Ca/P sedimentation method, liposome method) of all hard fibre cells and estrogen response element, cell all breaks, and the result is unsuitable for " external " and analyzes.
Table 1
Chemical compounds I a (Mol/L) cell * 10 -4
Contrast 12.3
2.10 -5?????????????0.1
10 -5??????????????2.8
5.10 -6?????????????7.0
10 -6?????????????10.0
Table 2
Chemical compounds I a (Mol/L) cell * 10 -4
Contrast 150
2.10 -5???????????????3
10 -5???????????????71
5.10 -6?????????????115
Table 3
Chemical compounds I a (Mol/L) cell * 10 -4
Contrast 7.6
2.10 -5?????????????0.1
10 -5??????????????5.4
5.10 -6?????????????6.3
10 -6??????????????7.6
Table 4
DNA DNA collagen collagen P value
(O: D.) (μ g) I type I type
(pg/ hole), (μ g/ μ g DNA) contrasts 4.85 ± 0.32 1.36 ± 0.06 47.82 ± 4.15 35.00 ± 1.41 chemical compounds 8.20 ± 0.23 1.97 ± 0.05 43.78 ± 5.23 22.00 ± 1.46 p＜0.005 I a, (1 μ M) chemical compound 6.90 ± 0.50 1.85 ± 0.24 38.01 ± 6.24 20.50 ± 0.61 p＜0.005 I a, (5 μ M) chemical compound 7.90 ± 1.46 1.96 ± 0.29 35.16 ± 2.44 18.00 ± 1.41 p＜0.005 I a, (10 μ M)

Claims (4)

1. a method that suppresses the aponeurosis fibromatosis comprises formula I compound or pharmaceutically acceptable salt thereof or solvate to the administration effective dose of this inhibition of need
Figure A9719191600021
R wherein 1And R 3Be independently hydrogen ,-CH 3,
Figure A9719191600022
Alkyl) or Wherein Ar is the optional phenyl that replaces;
R 2Be selected from pyrrolidine, hexamethyleneimino and piperidino.
2. the process of claim 1 wherein that described mammal is the people.
3. the method for claim 2, wherein said chemical compound is its hydrochlorate.
4. the method for claim 3, wherein said chemical compound is Or its hydrochlorate.
CN97191916A 1996-01-29 1997-01-27 Method of inhibiting musculoaponeurotic fibromatoses (desmoid tumors) Pending CN1209746A (en)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US1077396P 1996-01-29 1996-01-29
US60/010,773 1996-01-29
GBGB9603148.9A GB9603148D0 (en) 1996-02-15 1996-02-15 Methods of inhibiting musculoaponeurotic fibromatoses (desmoid tumors)
GB9603148.9 1996-02-15

Publications (1)

Publication Number Publication Date
CN1209746A true CN1209746A (en) 1999-03-03

Family

ID=26308707

Family Applications (1)

Application Number Title Priority Date Filing Date
CN97191916A Pending CN1209746A (en) 1996-01-29 1997-01-27 Method of inhibiting musculoaponeurotic fibromatoses (desmoid tumors)

Country Status (11)

Country Link
EP (1) EP0907361A4 (en)
JP (1) JP2000503995A (en)
KR (1) KR19990082055A (en)
CN (1) CN1209746A (en)
AU (1) AU707675B2 (en)
CZ (1) CZ235398A3 (en)
EA (1) EA199800678A1 (en)
IL (1) IL125523A0 (en)
NO (1) NO983452D0 (en)
PL (1) PL328127A1 (en)
WO (1) WO1997026878A1 (en)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU2135195C1 (en) * 1998-12-17 1999-08-27 Овсюк Валерий Григорьевич Method for producing material containing prostate cells and treating the cases of uterine fibromatosis, chronic prostatitis and male sexual function disorders by applying transplantation method
AU781168B2 (en) * 2001-01-26 2005-05-12 Pfizer Products Inc. Method of treating certain cancers using an estrogen agonist/antagonist

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
TW366342B (en) * 1992-07-28 1999-08-11 Lilly Co Eli The use of 2-phenyl-3-aroylbenzothiophenes in inhibiting bone loss
US5552162A (en) * 1993-02-09 1996-09-03 Arch Development Corporation Method for improvement of scar size and appearance
US5457116A (en) * 1993-10-15 1995-10-10 Eli Lilly And Company Methods of inhibiting uterine fibrosis
IL111287A0 (en) * 1993-10-15 1994-12-29 Lilly Co Eli Methods for treating resistant neoplasms
US5574047A (en) * 1993-12-21 1996-11-12 Eli Lilly And Company Methods of inhibiting imperfect tissue repair

Also Published As

Publication number Publication date
PL328127A1 (en) 1999-01-18
KR19990082055A (en) 1999-11-15
JP2000503995A (en) 2000-04-04
EP0907361A4 (en) 2002-05-02
IL125523A0 (en) 1999-03-12
NO983452L (en) 1998-07-27
AU2124097A (en) 1997-08-20
NO983452D0 (en) 1998-07-27
AU707675B2 (en) 1999-07-15
WO1997026878A1 (en) 1997-07-31
CZ235398A3 (en) 1999-02-17
EA199800678A1 (en) 1999-02-25
EP0907361A1 (en) 1999-04-14

Similar Documents

Publication Publication Date Title
CN1049335C (en) Methods for inhibiting endometriosis
CN1043608C (en) Improvements in or relating to benzothiophenes
CN1072931C (en) Methods of inhibiting cell-cell adhesion
CN1119530A (en) Methods of use for inhibiting bone loss and lowering serum cholesterol
US20050113307A1 (en) Methods of treating cancer and the pain associated therewith using endothelin antagonists
CN1058390C (en) Methods for exhibiting angiogenisis and angiogenic diseases
CN1108255A (en) Methods of inhibiting the symtoms of premenstrual syndrome/late luteal phase dysphoric disorder
CN1107156A (en) Methods of inhibiting uterine fiberosis
CN1107705A (en) Method for increasing libido in post-menopausal women
CN1107332A (en) Methods of inhibiting CNS problems in postmenopausal women
CN1107702A (en) Methods of inhibiting ovarian dysgenesis delayed puberty, or sexual infantilism
CN1108099A (en) Methods of inhibiting sexual precocity
CN1108100A (en) Methods of inhibiting breast disorders
CN1108103A (en) Methods of inhibiting turner&#39;s syndrome
CN1278812A (en) 2-aryl-3-aroylbenzo [b] thiophenes for treatment of estrin deficiency
CN1209746A (en) Method of inhibiting musculoaponeurotic fibromatoses (desmoid tumors)
EP1347751B1 (en) Methods of treating bone cancer and the pain associated therewith using endothelin antagonists
CZ106098A3 (en) Use of benzothiophene for preparing pharmaceutical preparation for inhibiting of 1 plasminogen activator inhibitor
AU688112B2 (en) Methods of inhibiting endometrial cancer
CN1108101A (en) Methods of inhibiting male infertility
US5843964A (en) Methods of inhibiting endometrial mitoses
US5670523A (en) Methods of inhibiting musculoaponeurotic fibromatoses (desmoid tumors)
CN1177295A (en) Method of inhibiting growth hormone effects
CN1193274A (en) Calcium-ion channel antagonists
CN1209745A (en) Method of inhibiting colon tumors

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C02 Deemed withdrawal of patent application after publication (patent law 2001)
WD01 Invention patent application deemed withdrawn after publication
REG Reference to a national code

Ref country code: HK

Ref legal event code: GR

Ref document number: 1045588

Country of ref document: HK