US20130012514A1 - Aminoalkyloxazole and aminoalkylthiazolecarboxylic acid amides as regeneration-promoting substances for sensory organs and post-mitotic tissue - Google Patents

Aminoalkyloxazole and aminoalkylthiazolecarboxylic acid amides as regeneration-promoting substances for sensory organs and post-mitotic tissue Download PDF

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US20130012514A1
US20130012514A1 US13/577,503 US201113577503A US2013012514A1 US 20130012514 A1 US20130012514 A1 US 20130012514A1 US 201113577503 A US201113577503 A US 201113577503A US 2013012514 A1 US2013012514 A1 US 2013012514A1
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cells
regeneration
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Holger Eickhoff
Hubert Löwenheim
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EMC Microcollections GmbH
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
    • C07D413/06—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P21/00—Drugs for disorders of the muscular or neuromuscular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/16—Otologicals
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D263/00—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings
    • C07D263/02—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings not condensed with other rings
    • C07D263/30—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
    • C07D263/34—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
    • C07D413/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
    • C07D417/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
    • C07D417/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a chain containing hetero atoms as chain links

Definitions

  • This disclosure relates to aminoalkyloxazole and aminoalkylthiazolecarboxylic acid amides that stimulate endogenous regeneration of terminally differentiated cells in highly specialized organs, tissues and sensory epithelia in mammals in situ.
  • the disclosure in particular relates to compounds with which a de novo formation of hair sensory cells in the organ of Corti can be obtained by inducing cell separation of supportive cells of the inner ear and hearing can be restored after hair cell loss.
  • the disclosure further relates to processes for preparing formulations of our compounds as pharmaceutical preparations and use thereof for producing pharmaceuticals for regenerative medicine.
  • hardness of hearing affects about 10% of the population of the industrialized nations. In Germany, there are estimated to be 16 million people who are hard of hearing, almost one fifth of the total population (ifo-Institut, 1986). Thus, hardness of hearing is not only the most frequent disorder of a sensory organ, but also one of the most frequent chronic disorders in general.
  • dedifferentiation plays the decisive role in regeneration of terminally differentiated tissue of amphibious animals.
  • other vertebrates have low regeneration abilities.
  • an extract can be obtained from amphibian tissues (for example, extremities) undergoing regeneration which is capable of inducing dedifferentiation even in mammalian cells (McGann et al., 2001).
  • amphibian tissues for example, extremities
  • this extract with appropriate stimulation, the dedifferentiation-based mechanism for regeneration of terminally differentiated cells can be transferred from amphibians to mammals (Odelberg, 2002).
  • regeneration extracts mentioned are “protein cocktails” and details with regard to their composition are not known.
  • R2 represents hydrogen or acyl and R1 and R3, which may be identical or different, represents a substituent selected from the group consisting of branched or straight-chain, substituted or unsubstituted alkyl groups, alkylcycloalkyl groups, alkylaryl groups, cycloalkyl groups, cycloalkylaryl groups, aryl groups and arylcycloalkyl groups which optionally contain heteroatoms, a pharmaceutically acceptable salt, a stereoisomer, a stereoisomer mixture, a tautomer or a prodrug compound, preferably a prodrug ester and a prodrug peptide, thereof.
  • FIG. 1 shows isolation and cultivation of stems cells from the organ of Corti of the mouse.
  • Stem cells from the inner ear can be obtained by individualizing the cells of the postnatal organ of Corti, isolating the stem cells still present and cultivating them under selective conditions in a suspension culture. Under these conditions, spheres are formed which express the stem cell marker Sox2 (A′) which is thought to have a decisive role in the pluripotency of embryonal stem cells and in the induction of pluripotent stem cells from differentiated cells (Takahashi and Yamanaka, 2006). Moreover, the nuclei in the spheres incorporate BrdU (A′′) which indicates the proliferation of the cells. (B) Several stem cell markers which can also be detected during ontogenesis in the organ of Corti are expressed in the spheres.
  • FIG. 2 shos in vivo and in vitro protein expressions profile of hair and supportive cell markers.
  • p27 Kip1 is expressed in all supportive cells of the organ of Corti. Neither inner nor outer sensory hair cells are p27 Kip1 -positive. It was possible to demonstrate p27 Kip1 expression within the epithelial islands (A′).
  • B GFAP labelling of supportive cells in the organ of Corti immediately below the nuclei of Deiter's cells and in the region of the inner phalangeal cells. The pattern of the filaments was also found in vitro (B′).
  • C E-Cadherin labels all supportive cells orientated laterally to the pillar cells. E-Cadherin is also located in the mebranes of the epithelial islands (C′).
  • sensory hair cell markers such as myosin VIIA (D), myosin VI (E) and calretinin (F) reliably label inner and outer sensory hair cells. All markers can also be detected in individual cells in the in vitro culture (D′, E′, F′).
  • the asterisks in each case mark the 3 outer sensory hair cells and the inner sensory hair cell (always on the right-hand side).
  • FIG. 3 shows relative number of cells expressing Sox2 in the screening assay.
  • FIG. 4 shows relative number of BrdU-positive cells in the screening assay.
  • the cells were incubated with BrdU for five hours.
  • FIG. 5 shows dose/activity analysis for N-cyclohexyl-2-[1-amino-2-(1H-indol-3-yl)ethyl]oxazole-4-carboxylic acid amide (3) (substance 33).
  • the dose-dependency of the induced BrdU effect was checked.
  • the BrdU incorporation could not be increased any further.
  • the control a population of cells from the in vitro culture with a comparable amount of DMSO without substance administration did not mediate any effect.
  • FIG. 6 shows incorporation of BrdU induced by N-cyclohexyl-2-[1-amino-2-(1H-indol-3-yl)ethyl]oxazole-4-carboxylic acid amide (3) (substance 33) in the organ culture model.
  • the BrdU-positive nuclei were at different stages of mitosis or had completed cell division (D, E, F).
  • FIG. 7 shows triple labelling after intracochlear administration of N-cyclohexyl-2-[1-amino-2-(1H-indol-3-yl)ethyl]oxazole-4-carboxylic acid amide (3) in vivo.
  • inner sensory hair cells labelled with myosin VI (marker for sensory hair cells) and BrdU (marker for cell division).
  • the cell nucleus is labelled with DAPI (nucleus staining).
  • Labelling with BrdU shows regeneration based on cell division.
  • Our low-molecular-weight compounds are able to induce corresponding cell biological changes such as dedifferentiation, proliferation and the subsequent terminal redifferentiation of cells of the normally post-mitotic tissue.
  • a precondition of the induction of this regeneration of the sensory hair cells is the suitable stimulation of the normally highly differentiated postmitotic auditory sensory epithelium.
  • Target cells are the supportive cells, which are directly adjacent to the damaged cells and which may serve as potential precursors for a reformation of sensory hair cells.
  • X represents O or S
  • Y represents C or N, where the two atoms must be different from one another
  • R2 represents hydrogen or acyl
  • R1 and R3 which may be identical or different, represent a substituent selected from the groups below: branched or straight-chain, substituted or unsubstituted alkyl groups, alkylcycloalkyl groups, alkylaryl groups, cycloalkyl groups, cycloalkylaryl groups, aryl groups and arylcycloalkyl groups which optionally contain heteroatoms, where R1 preferably represents (1H-indol-3-yl)ethyl and R3 preferably represents cyclohexyl.
  • aminoalkyloxazole and aminoalkylthiazolecarboxylic acid amides correspond to the formulae (3) to (8) below.
  • compositions of the formulae (1) to (8) include pharmaceutically acceptable salts (preferably non-toxic and physiologically tolerated salts), the stereoisomers, stereoisomer mixtures, all tautomers, prodrug compounds, preferably prodrug esters or prodrug peptides, and mixtures of the compounds of the formulae (1) to (8).
  • the concentration range for effective activity with regard to proliferation was defined to be in the range of from 0.1 ⁇ M to 100 ⁇ M, preferably from 1 ⁇ M to 3 ⁇ M, of substance in the cell culture medium.
  • Synthesis of the compounds is carried out in a number of steps. Detailed synthesis procedures can be found in Working Example A.
  • the starting materials for preparing the compounds are known and can be obtained from the specialist trade or be prepared by known procedures.
  • One possible route is the synthesis of the compounds on a solid phase starting with an amino acid amide.
  • the amino acid the side chain of which represents substituent R1
  • the amino acid thioamide is immobilized on a polymeric support having an activated carbonic ester group, and its acid group is then amidated.
  • the cyclization of the amino acid amide to the oxazole or of the amino acid thioamide to the thiazole is carried out using bromopyruvic acid and N,N-dimethylaniline. If ethyl bromopyruvate is employed, the ester group formed has to be hydrolyzed afterwards.
  • the amino acid amides are converted prior to the cyclization into the corresponding thioamides using Lawesson's reagent.
  • the amino acid oxazole/thiazole-carboxylic acids obtained are then reacted with an amine representing the substituent R3.
  • the amino acid oxazole/thiazolecarboxylic acid amides are cleaved with acid from the synthesis resin, followed by chromatographic purification.
  • the synthesis of the compounds is carried out in solution.
  • the synthesis takes place using protective group chemistry starting with a Boc-protected amino acid having the side chain R1.
  • the steps of the synthesis correspond to those described for the solid phase.
  • the Boc protective group is removed with acid, followed by chromatographic purification.
  • Both preparation processes permit the synthesis both of racemic and of enantiomerically pure compounds.
  • the compounds are obtained either in free form or as a salt, provided salt-forming groups are present.
  • Preferred salts of the compounds are pharmaceutically acceptable salts. Examples of such salts are salts of inorganic or organic acids, salts of inorganic or organic bases and salts of basic or acidic amino acids.
  • the compounds can also be modified as prodrug compounds, preferably as prodrug esters or prodrug peptides, or the like.
  • cell penetration-enhancing molecules such as, for example, biotin or maleimidopropionic acid, optionally via suitable spacer molecules, to the primary amino group, or by acylation of this amino group, corresponding to substituent R2, it is possible to improve the bioavailability and thus the efficacy of the compounds.
  • compositions for treating disorders in mammals associated with damaged postmitotic tissues, in particular for treating inner ear hardness of hearing in mammals caused by damage and loss of sensory hair cells in the organ of Corti.
  • These preparations comprising at least one active compound alone or in combination, optionally comprise further pharmaceutically suitable auxiliaries and additives such as, for example, carrier substances, preservatives, stabilizers, emulsifiers, detergents, solvents, solubilizers, salts for regulating the osmotic pressure and buffer salts.
  • auxiliaries and additives such as, for example, carrier substances, preservatives, stabilizers, emulsifiers, detergents, solvents, solubilizers, salts for regulating the osmotic pressure and buffer salts.
  • they may comprise further therapeutically relevant active compounds, adjuvants and also regeneration-promoting substances such as, for example, growth factors or anti-inflammatory agents.
  • Pharmaceutically suitable materials are the compounds known to be suitable for use in the field of pharmacy and food technology and related fields, in particular those listed in relevant pharmacopeias, whose properties do not exclude them from physiological administration.
  • Suitable administration forms of the pharmaceutical preparations comprising at least one active compound of formula (1) or (2) can be, for example, solutions, suspensions, sprays, gels, hydrogels, lotions, emulsions, pastes, ointments or creams.
  • the pharmaceutical preparations are prepared in a customary manner by known processes as described in relevant pharmacopeias, for example, by mixing, granulation or layering methods.
  • the pharmaceutical preparations may additionally be sterilized.
  • aminoalkyloxazole and aminoalkylthiazole-carboxylic acid amides as regeneration-promoting active compounds for treating, in humans and animals, disorders associated with damaged postmitotic tissues, preferably for recovery of hearing after loss or damage of sensory hair cells in the inner ear.
  • a person or an animal in need of such a treatment is administered a therapeutically effective amount of a regeneration-promoting preparation comprising at least one compound of the formula (1) or (2).
  • the active compound or the pharmaceutical preparation is applied directly or indirectly (including systemically), preferably locally, directly to/onto the damaged tissue structures.
  • Administration onto or into the inner ear takes place, for example, transtympanally by injection into the middle ear, by application onto the round or oval window of the inner ear or by injection into the inner ear.
  • Various systems for active compound administration may be employed.
  • the compounds and pharmaceutical preparations may be employed on their own, in combination with other of our compounds or in combination with one or more active compounds relevant for the respective therapeutic indication described of the compounds. There are no restrictions with regard to the sequence of administration. Our compounds may be administered simultaneously with, before or after the other active compounds, as a separate pharmaceutical or as a combination preparation, and by the same or by different administration routes.
  • the exact therapeutically effective amount for the treatment of inner ear hardness of hearing in a subject depends on various factors, inter alia on the extent of the tissue damage, on the height, stature, age and state of health of the patient, on the administration route and the administration form, the compound actually employed and, if appropriate, other pharmaceuticals used. Thus, at the current point in time, it is not expedient to specify the exact amount. In principle, however, repeated administration of the pharmaceutical preparations over a period of up to 8 weeks at intervals of from one to seven days to continuous administration using “sustained release” systems provided with a mechanism for the metered sustained release of active compounds may be assumed.
  • the amount of active compound employed should be in the range of from 0.5 ⁇ g to 1.0 mg per inner ear and administration.
  • Regeneration biologically relevant compounds capable of inducing corresponding cell biological changes such as dedifferentiation, proliferation or terminal redifferentiation and their formulations represent a novel form of active compounds since the therapeutic concept is completely new.
  • these compounds are suitable for use as pharmaceutically active compounds for the causal therapeutic treatment of sensorineural hearing loss on a regeneration biological basis.
  • this therapeutic approach is superior to all other methods discussed to date, such as gene therapy or stem cell transplantation.
  • gene therapy or stem cell transplantation In the in vitro and in vivo models examined to date, no negative effects were observed.
  • N-Boc-D-Tryptophan (9) (1 equiv.; 2.75 mmol; 1.00 g), hydroxybenzotriazole ammonium salt (2 equiv.; 5.50 mmol; 924 mg) and diisopropylcarbodiimide (DIC) (1.1 equiv.; 3.03 mmol; 382 mg; 454 ⁇ l) were dissolved in dimethylformamide (DMF) which had been dried over molecular sieves, and the solution was stirred at room temperature for 18 h. The solvent was then removed on a rotary evaporator under reduced pressure, and the residue was taken up in ethyl acetate.
  • DMF dimethylformamide
  • Chloro-(2′-chloro)trityl polystyrene resin (12) (Rapp Polymere H10033; loading 1.31 mmol/g) (1 equiv.; 655 ⁇ mol; 500 mg) was washed twice with DMF (over molecular sieve), a solution of the amino acid amide hydrochloride (13) (2 equiv.; 1.31 mmol) and diisopropylethylamine (DIPEA) (5 equiv.; 3.28 mmol; 424 mg) in DMF was added and the mixture was shaken at room temperature for 18 h. To cap the resin, methanol (MeOH) was then added to the suspension and the mixture was shaken at room temperature for another 15 min. The reaction solution was filtered off with suction, the loaded resin (14) was washed (5 ⁇ DMF, 3 ⁇ each MeOH, tetrahydrofuran (THF), diethyl ether) and dried under oil pump vacuum.
  • Ethyl bromopyruvate (5 equiv.; 3.28 mmol; 640 mg; 412 ⁇ l) and N,N-dimethylaniline (10 equiv.; 6.55 mmol; 794 mg; 832 ⁇ l) were dissolved in dioxane (5 ml) and added to the amino acid amide resin (14) (1 equiv.; 655 ⁇ mol; 500 mg).
  • the suspension was shaken at room temperature for 16 h and then warmed at 60° C. for 2 h.
  • the reaction solution was filtered off with suction and the resin was washed with dry solvents (5 ⁇ DMF, 5 ⁇ DCM).
  • reaction product (17) After a test cleavage using 5% TFA in DCM at room temperature for 1 h, identity and purity of reaction product (17) were checked by chromatography and mass spectrometry.
  • amino acid oxazolecarboxylic acid ethyl ester resin (15) or amino acid thiazolecarboxylic acid ethyl ester resin (17) (1 equiv.; 655 ⁇ mol; 500 mg) was pre-swollen in THF (2.5 ml), and a solution of lithium hydroxide monohydrate (5 equiv.; 3.28 mmol; 137 mg) in water (1.25 ml) and MeOH (1.25 ml) was added.
  • reaction solution was filtered off with suction, the resin (18) or (19) was washed (3 ⁇ each water, water/DMF 1:1, DMF, dioxane, THF, DCM, diethyl ether) and dried under reduced pressure in a desiccator.
  • the product (20) or (21) was then cleaved from the resin at room temperature using a solution of 5% TFA in DCM for 1 h.
  • Lawesson's reagent (0.75 equiv.; 750 ⁇ mol; 305 mg) was added to a solution of the Boc-amino acid amide (22) (1 equiv.; 1.00 mmol) in dry DME (7.5 ml).
  • the reaction solution was stirred at room temperature for 16 h and the solvent was then removed by distillation on a rotary evaporator under reduced pressure.
  • the residue was taken up in ethyl acetate (30 ml) and stirred vigorously with 10% strength sodium bicarbonate solution (15 ml) for 30 min. After separation of the phases, the aqueous phase was extracted twice with ethyl acetate.
  • the combined organic phases were washed three times with 10% strength sodium bicarbonate solution and then dried over sodium sulfate. After filtration, the solvent was distilled off and the product (23) was dried under oil pump vacuum.
  • the crude products can be recrystallized from ethyl acetate or ethyl acetate/petroleum ether or purified by flash chromatography on silica gel.
  • reaction solution was evaporated and the crude product was chromatographed on silica gel using a DCM/MeOH gradient.
  • the amide was stirred in 25% TFA in DCM at room temperature for 1 h and then evaporated. Repeatedly, heptane was added to the crude product and removed by distillation again on a rotary evaporator under reduced pressure. The product (25) was then lyophilized from tBuOH/water 4:1.
  • FIG. 1A By labelling on the protein level ( FIG. 1A ) and demonstrating a plurality of stem cell markers on the mRNA level ( FIG. 1B ), it was shown that the otospheres formed under these culture conditions are in a dedifferentiated state which corresponds to an earlier state during ontogenesis in the organ of Corti. In addition, it was possible to demonstrate cell divisions in the otospheres.
  • supportive cells are the potential precursors for a regeneration of sensory hair cells and therefore the actual target cells for the induction of sensory hair cell regeneration.
  • the cultivated otospheres were differentiated in a second step under adherent culture conditions on an ornithine/fibronectine surface to give monolayer epithelium islands relatively similar to the original sensory epithelium.
  • FIG. 2 shows a comparison of the situation in the native organ (in vivo) and in the differentiated epithelium islands of the cell culture (in vitro) using these markers.
  • Sox2 is a stem cell marker, which is also expressed during ontogenesis of the organ of Corti. It is thought to have an important role in the pluripotency of embryonal stem cells and during induction of pluripotent stem cells from differentiated cells (Takahashi and Yamanaka, 2006). Accordingly, Sox2 is an important marker, in particular in the context of an induced dedifferentiation/reprogramming of cells.
  • BrdU was then added to the culture medium simultaneously with the compounds (5 ⁇ M), and after 4 days the proliferation was quantified by BrdU incorporation.
  • the regeneration biological effect of the compounds was likewise demonstrated in the in vivo model of the adult guinea pig.
  • the compound N-cyclohexyl-2-[1-amino-2-(1H-indol-3-yl)ethyl]oxazole-4-carboxylic acid amide (3) was then continuously administered locally from a miniosmotic pump (Alzet®), subcutaneously implanted, via choleostomy directly into the scala tympani of the cochlea.
  • the dosage of the substance in the pump was correspondingly higher at 200 ⁇ M, taking into account an expected dilution effect in the perilymph.
  • BrdU was administered via the drinking water or the miniosmotic pump to label proliferating cells in the organ of Corti.
  • BrdU-labelled supportive cells and sensory hair cells were preferably demonstrated in the regions of the organ of Corti damaged by acoustic trauma. In control organs of the opposite side with/without acoustic damage and without substance administration, no BrdU-labelled sensory hair cells and, altogether, in accordance with the known findings on spontaneous proliferation, only few BrdU-labelled cells were found.

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US13/577,503 2010-02-08 2011-02-04 Aminoalkyloxazole and aminoalkylthiazolecarboxylic acid amides as regeneration-promoting substances for sensory organs and post-mitotic tissue Abandoned US20130012514A1 (en)

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DE102010007281A DE102010007281A1 (de) 2010-02-08 2010-02-08 Neue Aminoalkyl-oxazol- und Aminoalkyl-thiazolcarbonsäureamide und ihre Anwendung zur Stimulation der endogenen situ Regeneration von Haarsinneszellen im Corti'schen Organ des Innenohres beim Säuger
DE102010007281.8 2010-02-08
PCT/EP2011/000502 WO2011095338A1 (de) 2010-02-08 2011-02-04 Neue aminoalkyl-oxazol- und aminoalkyl-thiazolcarbonsäureamide als regenerationsfördernde substanzen für sinnesorgane und postmitotische gewebe

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EP2871181B1 (en) 2013-11-12 2016-10-05 Acousia Therapeutics GmbH Novel Compounds for Regeneration of Terminally-Differentiated Cells and tissues
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Family Cites Families (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5378803A (en) * 1987-12-11 1995-01-03 Sterling Winthrop Inc. Azole-fused peptides and processes for preparation thereof
DE19807426A1 (de) * 1998-02-23 1999-10-14 Otogene Biotechnologische Fors Verfahren zur Behandlung von Erkrankungen oder Störungen des Innenohrs
EP1126855B1 (en) * 1998-09-25 2007-05-09 Cephalon, Inc. Use of fused pyrrolocarbazoles for preventing/treating damage to sensory hair cells and cochlear neurons
TW502019B (en) * 1999-03-26 2002-09-11 Cocensys Inc Aryl substituted pyrazoles, imidazoles, oxazoles, thiazoles and pyrroles, and the use thereof
CN1441841B (zh) * 2000-07-11 2013-03-20 桑得医药品公司 对内耳的细胞再生和分化的刺激
UA89035C2 (ru) * 2003-12-03 2009-12-25 Лео Фарма А/С Эфиры гидроксамовых кислот и их фармацевтическое применение
JP2008502595A (ja) * 2004-03-31 2008-01-31 エグゼリクシス， インコーポレイテッド 未分化リンパ腫キナーゼモジュレータおよびその使用方法
JP5182088B2 (ja) * 2006-04-19 2013-04-10 アステラス製薬株式会社 アゾールカルボキサミド誘導体
GB0614538D0 (en) * 2006-07-21 2006-08-30 Univ Cambridge Tech Therapeutic Compounds And Their Use

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
Database Registry Chemical Abstracts Service, Columbus, Ohio, Accession No. RN 1023141-85-6, Entered STN: 28 May 2008. *
Database Registry Chemical Abstracts Service, Columbus, Ohio, Accession No. RN 1240216-68-5, Entered STN: 08 Sep 2010. *
Ito, Nobuyuki. Cancer Science 94(1), (2003) 3-8. *

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CN102947281A (zh) 2013-02-27
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DE102010007281A1 (de) 2011-08-11
WO2011095338A1 (de) 2011-08-11
CN102947281B (zh) 2015-11-25
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JP2013518917A (ja) 2013-05-23

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