CN1933840B - Use of a phenothiazine derivative for preventing and/or treating hearing loss - Google Patents
Use of a phenothiazine derivative for preventing and/or treating hearing loss Download PDFInfo
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Abstract
The invention concerns the use of a phenothiazine derivative of formula (I), wherein: R represents the hydrogen atom, an alkyl radical, arylalkyl, or -C(O)R', for preparing a medicine for preventing and/or treating hearing loss.
Description
The present invention relates to have calpain suppresses active and catches the purposes that the active phenothiazine derivative of active oxygen classification (ROS) is used to prepare the medicine that prevents and/or treats auditory dysesthesia.The present invention also relates to comprise the product of at least a such chemical compound and the pharmaceutical composition that contains this chemical compound.
In many reasons of auditory dysesthesia, can mention especially that the using of disease, heritability reason, damage, tumor, medicine, medicine such as some antibiotic, anticarcinogen, non-steroidal anti-inflammatory agent, diuretic, ulcer medicine or anticonvulsant as meningitis or otitis, long term exposure are in aromatic organic solvent such as toluene or dimethylbenzene, old and be exposed to noise.Presbyacusis (deafness relevant with the age), long term exposure are the main cause of auditory dysesthesia in using of noise and medicine.
Currently known is some antibiotic of aminoglycoside, as the gentamycin and the tobramycin that are used for the treatment of severe infections can cause the cochlea induced deafness.The toxicity of aminoglycoside (aminoglycoside such as amikacin, dibekacin, gentamycin, isepamicin, netilmicin, spectinomycin, tobramycin) shows by the hearing impairment at altofrequency place at first, and is not recognized by the patient at first.The patient only little by little is subjected to the puzzlement of this damage.Unfortunately it often is irreversible.
Noise around us has reduced our audition.When the hair cell that transmits sound to internal ear was damaged and no longer orders acoustic nerve to brain transmission electric pulse, the auditory dysesthesia that is caused by noise had just occurred.
The size of volume and the persistent period of exposure are two principal elements that influence auditory dysesthesia.Though interpersonal different, but still reported to have more deterministic facts to the reaction that is exposed to noise.Research shows that long term exposure is in 85 decibels (dB) or higher, through causing nonvolatil auditory dysesthesia after a while.
The statistics of Europe and North America shows that 8% to 10% population suffers from cochlea sexually transmitted disease (STD) reason (deaf, tinnitus).Consider the sound levels that discotheque, high-tech dance music meeting (techno concert) and walkman produce, producing the deafness and the tinnitus victim in a whole generation.So current relevant with the age problem (presbyacusis) that starts from about 60 years old can take place more early, promptly about 35-40 year.
The problem of audition pathology is that the overwhelming majority in them is because the loss of the neurocyte of hair cell and internal ear (or cochlea) causes.These cells except in uterus period of development basically, do not have the ability of self renewal after their last differential period.
Even fading of the sensitivity of the ear relevant with different cochlea pathology and neural aspect function is in the current scope that still surmounts any treatment.
So theme of the present invention is the purposes that the Hete rocyclic derivatives of the formula (I) that exists corresponding to any combining form with diastereomeric form or these forms is used to prepare the medicine that prevents and/or treats auditory dysesthesia,
Wherein R represents hydrogen atom, (C
1-C
6) alkyl, aryl alkyl or-C (O) R ' base, wherein R ' represents Heterocyclylalkyl, (C
1-C
6) alkyl, aryl or aralkyl;
Alkyl, aryl or Heterocyclylalkyl are optional to be selected from (C
1-C
6) alkyl, hydroxyl, (C
1-C
6) alkoxyl, nitro, cyano group, halogen or-NR
1R
2One or more identical or different substituent group replace; R
1And R
2Represent hydrogen atom or (C independently
1-C
6) alkyl, or R
1And R
2The nitrogen-atoms that is connected with them forms optional substituted heterocycle.
In this application, the one group of partial damage that relates to tissue and organ that causes by exterior materials of " wound " expression.Under the situation of audition wound, this exterior materials mainly is a noise.
(C
1-C
6) alkyl represents to comprise the alkyl group of the straight or branched of 1 to 6 carbon atom, for example methyl, ethyl, propyl group, isopropyl, butyl, isobutyl group, sec-butyl and the tert-butyl group, amyl group, neopentyl, isopentyl, hexyl, isohesyl.(C
1-C
6) alkoxyl is corresponding to above-mentioned alkyl group such as methoxyl group, ethyoxyl, propoxyl group or isopropoxy, but also comprise straight chain, the second month in a season or tert-butoxy.Alkyl-carbonyl can be corresponding to above-mentioned alkyl group such as methyl carbonyl, ethyl carbonyl, propyl group carbonyl.Halogen is represented fluorine, chlorine, bromine or iodine atom.
Aryl represents to comprise the carbocyclic ring or the heterocyclic ring system of at least one aromatic ring, and when at least one of the ring of forming system comprised hetero atom (O, N or S), this system was referred to as heterocycle.The example of the isocyclic aryl that can mention has phenyl or naphthyl.The example of the heterocyclic aryl that can mention (or heteroaryl) has thienyl, furyl, pyrrole radicals, imidazole radicals, pyrazolyl, isothiazolyl, thiazolyl, different
The azoles base,
Azoles base, pyridine radicals, pyrazinyl, pyrimidine radicals, benzothienyl, benzofuranyl and indyl.
Term heterocycle (or Heterocyclylalkyl) is preferably represented single or two ring fillings or unsaturated heterocycle, comprises 1 to 5 hetero atom that is selected from O, S and N.Nitrogen-atoms can be chosen the group that is selected from alkyl, aryl, aralkyl and alkyl-carbonyl wantonly and replace.As the example of saturated heterocyclic, can mention: oxolane, Pentamethylene oxide., oxetanes, oxepane, Tetramethylene sulfide, tetrahydric thiapyran, Thietane (thietane), pyrrolidine, piperidines, azetidine, 1,3-two
Alkane, 1,3-dioxolanes, 1,3-dithiolane, 1,3-dithiane, 1, the 3-oxathiolane (1,3-oxathiolane), 1,3-
Azoles alkane, 1,3-imidazolidine or 1,3-thiazoles alkane.As the example of unsaturated heterocycle, can mention: dihydro-thiophene, dihydrofuran, pyrrolin, glyoxalidine, pyrazoline, dihydropyridine, indoline.
Aryl alkyl (or aralkyl) refers to such group, and wherein aryl and alkyl are distinguished as defined above, as benzyl, phenethyl or naphthyl methyl.
At formula-NR
1R
2The situation of group under, R wherein
1And R
2The nitrogen-atoms that is connected with them forms can choose substituted heterocycle wantonly, this heterocycle preferably saturated and comprise 4 to 7 yuan of rings and comprise the nitrogen-atoms that existed at 1 to 3 interior hetero atom, extra hetero atom is independently selected from O, N and S atom.Described heterocycle can be as azetidine, pyrrolidine, piperidines, piperazine, morpholine or thiomorpholine ring.One or more identical or different substituent groups that described heterocycle can be selected from hydroxyl, alkyl, aryl, aralkyl or alkoxy base or halogen atom replace.
More particularly, theme of the present invention is a purposes as defined above, it is characterized in that R representative-C (O) R ' and preferred R ' represent alkyl group.
Most preferably, chemical compound (I) is characterised in that R representative-C (O)-CH
3Latter's chemical compound is called as chemical compound (1) hereinafter.
More particularly, theme of the present invention is a purposes as defined above, it is characterized in that R represents hydrogen.
Equally most preferably, chemical compound (I) has the structure of following formula as defined above
It more particularly is one of following formula
Equally most preferably, chemical compound (I) has the structure of following formula as defined above
And more particularly
This chemical compound is the protective agent with antioxidation and the effect of anticalcium activation neutral protease as defined above, is described among the application WO01/32654.
So theme of the present invention also is the purposes of formula (I) chemical compound as defined above, be used to relate to the pretreat or the back treatment of the cause of auditory dysesthesia.
Theme of the present invention also is the purposes of formula (I) chemical compound as defined above, is used to prepare the medicine that prevents and/or treats auditory dysesthesia behind another kind of medicament administration.Preferably, another kind of medicine is antibiotic such as gentamycin, anticarcinogen such as cisplatin, non-steroidal anti-inflammatory agent such as salicyclic acid derivatives or ibuprofen, diuretic such as furosemide, antiulcerative such as cimetidine or omeprazole, anticonvulsant such as carbamazepine or valproic acid.Most preferably, other medicine is an antibiotic, and more particularly is gentamycin.
Theme of the present invention also is the purposes of formula (I) chemical compound as defined above, is used to prepare the medicine that prevents and/or treats auditory dysesthesia behind presbyacusis.
Theme of the present invention also is the purposes of formula (I) chemical compound as defined above, is used to prepare the medicine that prevents and/or treats auditory dysesthesia after the audition wound.
According to chemical compound of the present invention can use separately or with at least a other materials with pharmaceutical active, the combinations of substances that preferably can prevent and/or treat auditory dysesthesia or prevent and/or treat any pathology relevant with auditory dysesthesia is used.
This chemical compound can with antioxidant, calpain inhibitor such as leupetin or Neurodur, peripheral vasodilator agent such as EGb 761
, nmda receptor agonist or antagonist, the kinase whose inhibitor peptides of c-Jun N-terminal such as D-JNK-1 make up.
Theme of the present invention also is a purposes as defined above, it is characterized in that it and at least a other combinations of substances with pharmaceutical active, described other materials are preferably and are selected from following material: the agonist of antioxidant, calpain inhibitor, peripheral vasodilator agent, nmda receptor or antagonist, the kinase whose inhibitor peptides of c-Jun N-terminal.
The chemical compound of formula (I) and preferred as defined above chemical compound (1), the dosage local application of 50 to the 500 μ M that can be comprised.Under the situation of partial cochlea internal therapy, it can be used with the dosage of 50 to 200 μ M.Under the situation of treatment, it can be used with the dosage of 200 to 500 μ M outside partial cochlea.Randomly with their the combination known substances on the pharmacology, the dosage of available common recommendation is used.
Chemical compound as defined above, and the optional material with pharmaceutical active that makes up with their can be by the route of administration of standard, and for example oral, intraperitoneal, subcutaneous or intravenous are used.They can be used by identical or different route of administration at the same time or separately.Preferably, chemical compound (I) is by the common technology local application in internal ear treatment as defined above, for example microtubular, through the syringe of injection of tympanum or be equipped with the conduit of the little gauze stripe shape of Silverstein meche.
Theme of the present invention also is (I) Hete rocyclic derivatives of formula as defined above and at least a product with material of therapeutic activity that comprises the combination in any of diastereomeric form or these forms, as combination product while, difference or the use of leap a period of time, to prevent and/or treat auditory dysesthesia.Preferably, theme of the present invention is a product as defined above, after drug administration, or after presbyacusis, or after the audition wound, be used to prevent and/or treat auditory dysesthesia.
At last, theme of the present invention is the product as defined above as medicine.
Under the situation of auditory dysesthesia behind the medicament administration, medicine is antibiotic preferably, and preferred gentamycin.
The optional material with pharmaceutically active that is combined is used by imagine the route of administration that is used for these materials usually in this treatment field.
Under the situation of the auditory dysesthesia that causes by the audition wound, using of chemical compound (I) as defined above can occur in a couple of days before the audition wound, and preferably before wound 2 to 3 days, and after wound 24 hours.Preferably, after wound, use in 7 days.Equally preferably, after wound, use in 2 hours.
So theme of the present invention is aforesaid purposes, it is characterized in that the chemical compound (1) of the combination in any of diastereomeric form or these forms, after wound, use in 7 hours, preferably after wound, use in 1 hour.
Experimental section provide be presented at the audition wound after chemical compound (1) at the treatment effectiveness aspect the functional rehabilitation.
Provide the following example so that above-mentioned method to be described, should not be understood as that to limit the scope of the invention.
Experimental section:
Pharmaceutical research
1) treats inductive ototoxicity with gentamycin
The chemical compound of using as co-therapy 1 is for the confirmation by the protective effect of the inductive hair cell loss of gentamycin
Gentamycin and other aminoglycoside medicaments have shown the damage and the auditory dysesthesia that can cause hair cell in the mankind.Brachydanio rerio is called as neuromast at the surface of their healths expression sensory organ.In these fishes, the neuromast hair cell can dye with DASPEI, and this dyeing reflects the quantity of hair cell.These hair cells are similar to the interior hair cell of people's ear on 26S Proteasome Structure and Function.
In Brachydanio rerio, induce the damage of internal hair cell by gentamycin.For the effect of test compounds (1) to the protection of the damage of the hair cell that caused by gentamycin, chemical compound (1) is used with gentamycin as the co-therapy medicine.Dyeing and quantitatively internal ear hair cell then.
Research is carried out in the fish of 5 ages in days, and the gentamycin with 1 μ g/ml under the situation that has or do not exist chemical compound (1) was cultivated 24 hours.Use single carrier (1%DMSO abreast; Positive control) contrasts.The negative contrast of fish with the gentamycin processing.
Carry out DASPEI (iodate 2,4-dimethyl-aminobenzene vinyl-N-ethylpyridine
) dyeing, so that show (n=5/group) hair cell in vivo.Use morphometric Analysis to carry out quantitatively with dyeing signal to hair cell.The DASPEI dyeing signal of positive control is defined as 100%.
The result is shown in Fig. 1 (with the painted hair cell percent of DASPEI.Positive control (Brachydanio rerio-1%DMSO); The effect (Brachydanio rerio-1%DMSO-gentamycin-chemical compound 1) of negative control (Brachydanio rerio-1%DMSO-gentamycin 1 μ g/ml) and this product.Experiment is carried out with every group of 5 animals).
The result of Fig. 1 shows:
The dyeing signal of-negative control is represented 31.5 ± 4.2% control signal, perhaps handles the hair cell loss of back 68.5 ± 4.9% with gentamycin.
The dyeing signal of-animal of handling with gentamycin and chemical compound 1 represent 65.2 ± 4.4% control signal, or with the protection of 48.7 ± 2.63% highly significants of the impaired hair cell of gentamycin processing.
2) the post-traumatic auditory dysesthesia of audition
This relates to after the auditory dysesthesia that is caused by the audition wound; the protective effect research of chemical compound in Cavia porcellus (1); this protective effect relates to the protection to the internal ear hair cell in one aspect before treatment and after the treatment, relate to the functional rehabilitation of audition on the other hand.By local approach " cochlea in and cochlea outer " administered compound (1) with the purposes of simulation in people's surgical clinical medicine.
Can be quantitative with the function and protecting that this new protective agent obtains with function test, this function test is the reading of animal audiogram.This audiogram produces by the activity that writes down acous potential compound action.Before the audition wound and write down audiogram afterwards.Analysis by scanning electron microscope is finished this performance data by research along the loss cell of cochlea spiral and the protective effect of chemical compound (1).
All experiments are all carried out in Cavia porcellus; Carry out each experiment according to similar scheme.
The general approach of experiment periods is as follows:
-with the 2%Rompun of animal with intramuscular injection
(3mg/kg)+Zoletil
Mixture (40mg/kg) is anaesthetized.This anesthesia has the advantage of rapid dissipation, and can keep a few hours by injecting 1/3rd of (per 2 hours) predose regularly.
-according to the placement of electrode and the micropump of Fig. 2.
Approach by dorsal part is near cochlea.After scalp being shaved off hair and cleaning, behind auricle, cut the otch of a 2cm.The parotid gland and the muscle that cover bulla tympanica are tiltedly put.After dry and cleaning, pierce through bone at its upper wall, expose nervus facialis below.Under the control of operating microscope (WILD M650), will contact placement in active electrode (the platinum lead of diameter 0.13mm wraps up with polytetrafluoroethylene) the importing bubble and with the film of fenestra cochleae then.After tin threshold value that record is measured, change hands the worker and bore the aperture that a diameter is 0.2mm at the end of the tympanic canal below fenestra cochleae.
Use second micro-manipulator, will be inserted in the cochlea by the glass pipet (vertical diameter is 0.1mm) that conduit links to each other with Micropump.The bulla tympanica that comprises recording electrode and perfusion pipet seals with the dentistry resin.
With Micropump slide into zoodermic below, with povidone iodine muscle and skin plane are carried out wiping, sew up and cover antibiotic solution (Rifocin 5P100 with absorbable suture
).
Second otch that overhead go up to cut about 1.5cm with fixed connector on the animal skull (Connectral, ref.:8/45-05.050.000).After scraping off periosteum, carefully drying should the zone, applies silver nitrate, covers with the cyanolite film then.
Will (effectively and reference) electrode slide into and subcutaneously combine with adapter.Then adapter is fixed on the skull with the dentistry resin.
Stimulating technology:
Use two Hewlett Packard synthesizers (HP3314 A and HP8904 A) to produce sonic stimulation, and in free found field, transmit by speaker (JBL075) apart from ear 10cm.Use 1/2 inch mike (4134 types, Bruel and Kjaer) and can be directly with decibel SPL (dB SPL, reference: 2.10-5Pa) read the measuring amplifier (2606 type) of sound levels, in artificial ear, carry out the calibration of sound system.In order to show acoustical signal, the output of measuring amplifier links to each other with oscillograph.Animal was exposed on 6kHz, 120dB SPL following 30 minutes.
Recording technique:
Use preamplifier and GRASS P511 K type difference amplifier, the cochlea reaction of writing down by the electrode of implanting cochlea via the adapter that is fixed in the animal head is amplified (increasing by 1000 times) and filtered (32Hz-3200Hz).Directly trace is presented at (Tektronix 513 types) on the oscillograph.This signal is averaged (256 acceptance values) reducing background noise, and be stored among the 486PC of 66 megahertzes (Hewlett-Packard-Vectra 05/65).The definition threshold criteria is for producing the essential dB SPL value of measurable reaction (＞2 μ V).Contact the audiogram that two electrodes placing can write down the reaction of cochlea and produce each ear with cochlea (fenestra cochleae).After the audition wound, write down tin threshold value of 20 minutes measurement every day, one month by a definite date.
The activatory participation of calpain after the audition wound
Cutting to the specific substrate fodrin of calpain is carried out quantitatively, to determine the activation of calpain after the audition wound.The fodrin of calpain cutting 240KD is to form the catabolite of 150KD.With to the special polyclonal antibody of 150KD segment and can identify that the conjugated protein antibody of anticalcium of hair cell carries out double labeling.Under confocal microscope, show fluorescence.
The molecule mechanism of hair cell death after the audition wound:
In order to determine the essence of cell death, use the TUNEL method that the dna segmentization of cells,cochlear has been carried out quantitatively.
The hair cell integrity determines after the audition wound
Use anti-cell pigment C antibody to determine the integrity of cells,cochlear by immunocytochemistry.In healthy cell, cytochrome C is positioned mitochondrion.After the audition wound, the cytochrome diffusion also is distributed in endochylema.
1 phase: the chemical compound of using as pretreat (1) is to auditory dysesthesia and the proof along the protective effect of the loss cell of cochlea spiral that caused by wound
Scheme-1 phase: the using in advance of chemical compound (1)
Be placed on the infiltration micro pump delivered chemical compound (1) under the skin.
Implant two days later at micropump, in cochlea, carry out the audition wound via conduit (perfusion in the cochlea) animal.
In order to determine dose response, this experiment is carried out in 7 animals, carries out in 30 animals then.
Infiltration micropump (flow velocity of 1 μ l/h, the volume of 200 μ l, 7 days persistent period of diffusion) via at preceding 2 days permanent implanted cochlea of wound directly applies to (perfusion in the cochlea) in the cochlea with chemical compound (1).This technology can determine chemical compound (1) when the dosage of 100 μ M to cause by wound along the protective effect of the loss cell of cochlea spiral and the functional restoration effect of audition.The dosage effect of product can determine to preserve the effective dose of 50% audition.
1 phase result
The chemical compound (1) of 100 μ M concentration.
Test by the audiogram measurement function.Protect at the post-traumatic auditory dysesthesia of audition with chemical compound 1.
The wound that 120dB continues 30 minutes is after five days, the effectiveness of the provable chemical compound of the reading of audiogram (1); When with the concentration of 100 μ M during in wound perfusion in preceding 2 days, this product can recover 100% audition (Fig. 3).
Morphology research: after the audition wound along the histology of the cell of cochlea spiral.The protective effect of chemical compound (1).
When electrophysiology evaluation (audiogram) finishes, after the audition wound 30 days, the cochlea of animal is taken a sample, and the preparation sample is used for electron micrograph.
With scanning microscope by ciliary plexus (ciliary tuft) being counted to determine the loss of cell.Amplify observation to seek qualitative data by the height that carries out Cole's base of a fruit (Corti) device surface.
After the wound that continues 30 minutes at 120dB, some among three rows of interior hair cell and outer hair cell are damaged.The effectiveness of the provable chemical compound of the histology of these cells (1).When preceding 2 days of wound is poured into 100 μ M, this product makes 100% interior hair cell and the most outer hair cell (Fig. 4: histological data that is protected; Picture A: audition wound; Photo B: chemical compound (1)+audition wound; Hair cell in the I=; The outer hair cell of O=).
The activatory participation of calpain after the audition wound.Suppress with chemical compound 1.
The activation of calpain is that the specific substrate fodrin by this enzyme is degraded into that 150KD is pulsating quantitatively to be determined.
In the cells,cochlear of contrast, never detect the cutting of fodrin.
After the audition wound because the activation of calpain, the 150KD immune marker that derives from the fodrin cutting outside on the cells,cochlear because with anti-FBDP antibody (" fodrin catabolite ", green fluorescence; Fig. 5 A: after the audition wound 48 hours) labelling but visible.The active activation of calpain is relevant with the loss of cells,cochlear after the audition wound, and this loss shows that by there not being the conjugated protein antibody of anticalcium described antibody can be identified complete hair cell.
The chemical compound (1) of topical application 100 μ M---a kind of calpain activation inhibitor antioxidant can prevent to be exposed in the cells,cochlear of audition wound calpain to the cutting of fodrin.There is not the green fluorescence labelling, so the degraded of the nothing of the specific substrate of calpain is relevant with the protection of cells,cochlear (with the conjugated protein antibody labeling of anticalcium; Fig. 5 B).
The molecule mechanism of hair cell death after the audition wound.The protection of chemical compound 1.
For the essence of the cells,cochlear death determining to cause, in the animal that suffers the audition wound, carry out the fragmentation (quantitative) of DNA by the TUNEL method by the audition wound.With the effect of this parameter testing chemical compound 1, participate in by this mechanism of cell death that apoptosis causes to determine it.
Do not observe any " the TUNEL positive " cell in the cochlea that in the control animal that is not exposed to noise, obtains.
The cochlea that is exposed to the audition wound has TUNEL positive cell nuclear (Fig. 6 A: after the audition wound 48 hours) in Cole's governor zone.Appraise and decide the last zone that is positioned at Cole's governor significantly.So these significant nuclears belong to hair cell rather than sustenticular cell.Many significant nuclears after the audition wound one hour be visible, and this is marked at the audition wound and is still visible after 4 days.So the cell death of sound hair cell produces via the apoptosis mechanism relevant with the fragmentation of DNA after the audition wound.
The chemical compound (1) of topical application 100 μ M---a kind of calpain activation inhibitor and antioxidant, labelling that can prevention sound capillary karyon (TUNEL method, Fig. 6 B: chemical compound (1)-after the audition wound 48 hours).Chemical compound (1) suppresses to pass through the cell death that apoptosis carries out by what the audition wound caused.
The loss of hair cell integrity after the audition wound.Protect with chemical compound (1).
In order to determine the loss of hair cell integrity after the audition wound, in the animal that suffers the audition wound, carry out the diffusion of cytochrome C, this diffusion uses anti-cell pigment C antibody to carry out by immunocytochemistry.
Test compound (1) the release of pair cell pigment C from the mitochondrion compartment to the endochylema compartment is to determine after this chemical compound (1) is to the audition wound effect in the integrity loss of hair cell.
In the cochlea that obtains in not being exposed to the control animal of noise, cytochrome C is positioned mitochondrion.
The cochlea that is exposed to the audition wound has shown the labelling (Fig. 7 A) of the cytochrome C that spreads and distribute in endochylema.
The chemical compound (1) of topical application 100 μ M---a kind of calpain activation inhibitor and antioxidant, can prevent to be exposed in the hair cell of audition wound cytochrome C from the diffusion of mitochondrion, so kept cell integrity (Fig. 7 B) to endochylema.
The dose-effect curve of chemical compound (1).
Recover 50% effective dose (ED of hearing loss with definition by change dosage
50), the protective effect that comes assessing compound (1).Eight groups of every group of 5 animals comprise those animals of only accepting artificial perilymphatic those animals and accepting 1,3,10,33,100 μ M chemical compounds (1),, have used 30 animals that is.
After the wound of 120dB continues 30 minutes five days, the reading of audiogram can determine that chemical compound (1) allows to recover 50% effective dose (ED of hearing loss after the wound
50=3.61 μ M) (Fig. 8).
2 phases: by handling chemical compound (1) that back cochlea outer filling uses to confirmation along the protective effect of the loss cell of cochlea spiral and auditory dysesthesia.After wound, chemical compound (1) makes and recovers determining of 50% audition required time.
Use behind the chemical compound (1):
Preliminary research shows: the cochlea otomy of carrying out after exposure has increased the weight of the wound effect of sound.From this on the one hand, implanted animal can not recover well as the animal that does not have to implant after the wound.In order to eliminate the wound relevant with the cochlea otomy, we have developed the noninvasive method that chemical compound is directly applied to fenestra cochleae (cochlea is outer).
Osmotic minipumps is implanted subcutaneous, after animal suffers the audition wound 30 minutes or 1,3,6,12 or 24 hour, osmotic minipumps was sent chemical compound (1) into cochlea (cochlea outer filling) by conduit.
In 30 animals, carry out this experiment.
Via osmotic minipumps (flow velocity 1 μ l/h, 200 μ l volumes, 7 days persistent period of diffusion), chemical compound (1) was applied to middle ear (cochlea outer filling) in 1,3,6,12 or 24 hour after preceding 48 hours of wound or wound.This Micropump is permanently implanted middle ear and product directly is diffused on the fenestra cochleae.
These experiments can be determined the drug holiday (DH) of chemical compound (1), promptly can give the maximum duration of chemical compound (1) chemical compound (1) demonstration simultaneously protective effect after wound.Also determine the 50% effective time (ET that recovers of loss audition after wound
50).This technology also allows to determine in the protective effect along the loss cell of spiral cochlea that chemical compound (1) caused wound under the dosage of 300 μ M in month after the audition wound.
2 phase results
Chemical compound (1) when 300 μ M concentration.
Functional experiment: audiogram.
After wound one hour, chemical compound (1) was kept 90% audition.Be determined to be between 6 to 7 hours giving time of 50% that chemical compound (1) meanwhile keeps audition after the wound.The treatment window of chemical compound (1) is 24 hours after the audition wound that continues 30 minutes at 120db.This means in this model after the audition wound first 24 hours chemical compound (1) be activated (consult Fig. 9: the audiogram that carried out in 10 days after the audition wound is measured).
Morphology research: month histology after the audition wound along the cell of cochlea spiral.
The cochlea outer filling of 300 μ M chemical compounds (1) of beginning in six hours was still protected most of hair cell in post-traumatic one month in audition after the audition wound.In fact, in zone by the audition trauma injuries, have only 32% interior hair cell and 18% outer hair cell not to exist, in contrast be, in zone by the audition trauma injuries, be exposed to noise but chemical compound of no use (1) treat have 86% interior hair cell and 62% outer hair cell not to exist to the side.
Claims (21)
1. the Hete rocyclic derivatives corresponding to formula (I) that exists with any combination of diastereomeric form or these forms is used to prepare the purposes of the medicine that prevents and/or treats auditory dysesthesia,
Wherein R represents hydrogen atom, (C
1-C
6) alkyl or-C (O) R ' base, wherein R ' representative (C
1-C
6) alkyl;
Alkyl is optional to be selected from (C
1-C
6) alkyl, hydroxyl, (C
1-C
6) alkoxyl, nitro, cyano group, halogen or-NR
1R
2One or more identical or different substituent group replace;
R
1And R
2Represent hydrogen atom or (C independently
1-C
6) alkyl.
2. according to the purposes of claim 1, it is characterized in that R representative-C (O) R '.
3. according to the purposes of claim 2, it is characterized in that R representative-C (O)-CH
3
4. according to the purposes of claim 1, it is characterized in that R represents hydrogen.
5. according to each purposes of claim 1-4, it is characterized in that chemical compound (I) has the structure of following formula
6. according to the purposes of claim 5, it is characterized in that chemical compound (I) has the structure of following formula
7. according to the purposes of claim 5, it is characterized in that chemical compound (I) has the structure of following formula
8. according to each purposes of claim 1 to 4, it is characterized in that chemical compound (I) has the structure of following formula
9. according to the purposes of claim 7, it is characterized in that chemical compound (I) has the structure of following formula
10. according to the purposes of claim 7, it is characterized in that chemical compound (I) has the structure of following formula
11., it is characterized in that derivant (I) uses with treatment in advance according to each purposes of claim 1-4.
12. according to each purposes of claim 1-4, it is characterized in that derivant (I) after treatment use.
13., be used to prepare the medicine that behind medicament administration, prevents and/or treats auditory dysesthesia according to each purposes of claim 1-4.
14., after the other drug that is selected from antibiotic, anticarcinogen, non-steroidal anti-inflammatory agent, diuretic, antiulcerative, anticonvulsant is used, use according to the purposes of claim 13.
15., after antibiotic administration, use according to the purposes of claim 14.
16., it is characterized in that antibiotic is a gentamycin according to the purposes of claim 15.
17., be used to prepare the medicine that behind presbyacusis, prevents and/or treats auditory dysesthesia according to each purposes of claim 1-4.
18., be used to prepare the medicine that after the audition wound, prevents and/or treats auditory dysesthesia according to each purposes of claim 1-4.
19., it is characterized in that chemical compound (I) uses in 7 hours after the audition wound according to the purposes of claim 18.
20., it is characterized in that chemical compound (I) uses in 1 hour after the audition wound according to the purposes of claim 19.
21. according to each purposes of claim 1-4, it is characterized in that chemical compound (I) and at least a other combinations of substances with pharmaceutical active, described other materials can prevent and/or treat auditory dysesthesia or prevent and/or treat any pathology relevant with auditory dysesthesia, and are selected from: the agonist of antioxidant, calpain inhibitor, peripheral vasodilator agent, nmda receptor or antagonist, the kinase whose inhibitor peptides of c-Jun N-terminal.
Applications Claiming Priority (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| FR0403203 | 2004-03-29 | ||
| FR0403203A FR2867979B1 (en) | 2004-03-29 | 2004-03-29 | NOVEL THERAPEUTIC APPLICATION OF A PHENOTHIAZINE DERIVATIVE |
| FR0406404 | 2004-06-14 | ||
| FR0406404 | 2004-06-14 | ||
| PCT/FR2005/000713 WO2005092345A1 (en) | 2004-03-29 | 2005-03-25 | Use of a phenothiazine derivative for preventing and/or treating hearing loss |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1933840A CN1933840A (en) | 2007-03-21 |
| CN1933840B true CN1933840B (en) | 2010-12-15 |
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ID=34947604
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| Application Number | Title | Priority Date | Filing Date |
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| CN2005800091391A Expired - Fee Related CN1933840B (en) | 2004-03-29 | 2005-03-25 | Use of a phenothiazine derivative for preventing and/or treating hearing loss |
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| CN (1) | CN1933840B (en) |
| FR (1) | FR2867979B1 (en) |
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| FR2898357B1 (en) * | 2006-03-07 | 2012-02-17 | Scras | AMIDINE DERIVATIVES AND THEIR APPLICATIONS AS A MEDICINAL PRODUCT |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1391571A (en) * | 1999-11-05 | 2003-01-15 | 科学研究和应用咨询公司 | Heterocyclic compounds and their use as medicines |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| FR2816509B1 (en) * | 2000-11-15 | 2004-02-06 | Sod Conseils Rech Applic | ASSOCIATION OF CALPAIN INHIBITORS AND OXYGEN REACTIVE TRAPS |
-
2004
- 2004-03-29 FR FR0403203A patent/FR2867979B1/en not_active Expired - Fee Related
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1391571A (en) * | 1999-11-05 | 2003-01-15 | 科学研究和应用咨询公司 | Heterocyclic compounds and their use as medicines |
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| 同上. |
| 权利要求9-12 |
| 说明书第1页1-10行,第2页5-6行,第2页17行至第4页9行,第7页1-4,23行,第8页5-8,15-21行,第11页2行至第12页16行,第18页17行至第28页第18行. |
| 说明书第1页1-27行,第4页1-12行,第6页11行至第7页12行,第11页28行至第12页2行,第12页14-25行. |
| 说明书第1页第1段,第7页12-16行. |
| 说明书第7页12-16行. |
Also Published As
| Publication number | Publication date |
|---|---|
| FR2867979B1 (en) | 2006-06-30 |
| FR2867979A1 (en) | 2005-09-30 |
| CN1933840A (en) | 2007-03-21 |
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