EP2370106A2 - Construction de principe actif et de peptide pour une administration extracellulaire - Google Patents

Construction de principe actif et de peptide pour une administration extracellulaire

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Publication number
EP2370106A2
EP2370106A2 EP09774835A EP09774835A EP2370106A2 EP 2370106 A2 EP2370106 A2 EP 2370106A2 EP 09774835 A EP09774835 A EP 09774835A EP 09774835 A EP09774835 A EP 09774835A EP 2370106 A2 EP2370106 A2 EP 2370106A2
Authority
EP
European Patent Office
Prior art keywords
active ingredient
peptide
construct according
peptide construct
construct
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.)
Withdrawn
Application number
EP09774835A
Other languages
German (de)
English (en)
Inventor
Gunter Fischer
Miroslav Malesevic
Frank Erdmann
Jan KÜHLING
Michael Bukrinsky
Stephanie Constant
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.)
Max Planck Gesellschaft zur Foerderung der Wissenschaften eV
Original Assignee
Max Planck Gesellschaft zur Foerderung der Wissenschaften eV
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Filing date
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Application filed by Max Planck Gesellschaft zur Foerderung der Wissenschaften eV filed Critical Max Planck Gesellschaft zur Foerderung der Wissenschaften eV
Publication of EP2370106A2 publication Critical patent/EP2370106A2/fr
Withdrawn legal-status Critical Current

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/001—Preparation for luminescence or biological staining
    • A61K49/0013—Luminescence
    • A61K49/0017—Fluorescence in vivo
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
    • A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
    • A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
    • A61K47/645—Polycationic or polyanionic oligopeptides, polypeptides or polyamino acids, e.g. polylysine, polyarginine, polyglutamic acid or peptide TAT
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
    • A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
    • A61K47/645—Polycationic or polyanionic oligopeptides, polypeptides or polyamino acids, e.g. polylysine, polyarginine, polyglutamic acid or peptide TAT
    • A61K47/6455—Polycationic oligopeptides, polypeptides or polyamino acids, e.g. for complexing nucleic acids

Definitions

  • the present invention relates to an active-peptide construct for extracellular enrichment, to a method for accumulating active substances in an extracellular space of a multicellular object, to the use of the active ingredient-peptide construct according to the invention for the production of a medicament and to a pharmaceutical composition comprising the active ingredient according to the invention.
  • peptide construct for extracellular enrichment, to a method for accumulating active substances in an extracellular space of a multicellular object, to the use of the active ingredient-peptide construct according to the invention for the production of a medicament and to a pharmaceutical composition comprising the active ingredient according to the invention.
  • active substances which are usually active pharmaceutical ingredients, usually exert their effect both inside and outside of biological cells, with the primary problem being that active substances which have their effect only within their own limits In addition, the problem arises that the same active substance can have a different effect inside and outside the cell, even before they pass through the cell membrane If the intracellular effect is to be achieved, since the transport to the cell usually involves the crossing of an extracellular space, the extracellular (secondary) effect must necessarily also be accepted.
  • Active ingredients can act extracellularly on molecules or structures.
  • biological molecules of the extracellular space can be, for example: enzymes, inhibitors, activators or receptors.
  • structures is meant, for example, the extracellular matrix formed from all the macromolecules located outside the plasma membrane of cells in tissues and organs.
  • the object of the present invention was therefore to find a way in which active substances can be administered to a multicellular object without the administered active ingredients being able to penetrate into the cell interior of the multicellular object.
  • the active ingredient-peptide construct according to the invention not only has the advantage that active ingredients can be used in a targeted manner in an extracellular space, it also offers the possibility of further disadvantages, which are associated with the use of certain active ingredients targeted by the use of specially composed peptides to compensate.
  • the activity of active ingredients which are sparingly soluble in water and therefore in most extracellular tissues can be improved by linking with peptides which additionally have a very high water solubility.
  • This also has the advantage that, in turn, the amount of active ingredient to be used can be reduced.
  • the object of the present invention is therefore achieved by a drug-peptide construct comprising an active ingredient A and a peptide B, wherein the construct has a negative net charge at pH 6 and wherein the active ingredient-peptide construct is free of a constituent C, which can walk through the membrane of a biological cell.
  • a drug-peptide construct is understood to mean any molecule which comprises an active compound A which is linked to at least one peptide B.
  • the connection between active ingredient A and peptide B can be carried out in any way known to those skilled in the art.
  • the active ingredient A and the peptide B of the active ingredient-peptide construct according to the invention are covalently linked directly to one another, but it is also preferred that the active ingredient A and the peptide B are linked to one another via a linker.
  • a molecule as a linker is characterized by the fact that by its nature no or only a negligible contribution to the function of the linker and serves only to bring the distance between a first part X of a total molecule which has one property to another part Y of a total molecule which has an identical or different property as X to a desired length.
  • diamines eg ethylenediamine or
  • Diaminopropane polyfunctional acids, polyethylene oxide, polypropylene oxide, amino acids, amino acid derivatives or ethylene glycol can be e.g. Form oligomers containing 2 to 10 or combinations of these monomeric units.
  • Linkers can also be composed of branched or unbranched alkanes.
  • linkers examples include e.g. the formation of amides from carboxylic acids and amines, secondary and / or tertiary amines from haloaliphatics and amines, the addition to double bonds, the formation of ethers or thioethers from halo-carboxylic acids and thiols, thioethers from thiols and maleimides, amide bonds from thioesters and 1,2-aminothiols, thioamide linkages of dithioesters and 1,2-aminothiols, thiazolidines of aldehydes and 1,2-aminothiols, oxazolidines of aldehydes / ketones and 1,2-aminoalcohols, imidazoles of aldehydes / ketones and 1 , 2-diamines (such as used in FIG. 3), of thiazoles of thioamides and alpha-halo-ketones
  • Isothiocyanato-ketones of oximes of amino-oxy-compounds and aldehydes, of oximes of amino-oxy-compounds and ketones, of hydrazones of hydrazines and aldehydes, of hydrazones of hydrazides and ketones and numerous others.
  • methods are particularly preferred which generate linkers with atomic distances between 4 and 40.
  • the construct according to the invention comprises a linker
  • the linker it is preferred if the linker has a chain of 4 to 40 C-C bonds.
  • the active ingredient-peptide construct may be both a compound of one or more active substances A of the same or different kind with one or more peptides B of identical or different nature. Further, in this case, the peptide (s) B and the drug (s) A may be linked in the same or different ways. Accordingly, the term “active ingredient-peptide construct” also means a construct of one or more active substances which are understood by the term “active ingredient A v and one or more peptides B which can be subsumed under the term” peptide B ".
  • the drug-peptide construct of the invention must continue to have a net negative charge at pH 6 to achieve the object of the invention.
  • the determination of the net charge of a peptide sequence can be carried out as a first approximation by calculation, as for example in WO 2003/097706, or more precisely and according to the invention by appropriate biochemical experiments such as isoelectric focusing (Encyclopedia of Biochemistry) or titration (according to Fresenius 1 Journal of Analytical Chemistry 274 (1975) 359-361) or according to Helvetica Chimica Acta. 91 (2008) 468-482).
  • the construct in order to ensure that the construct according to the invention remains in the extracellular space, the construct must furthermore not comprise any constituent C which is capable of traversing the membrane of a biological cell.
  • a "constituent C" is understood as meaning an additional constituent which differs from the constituents A (active substance) and B (peptide) of the construct or is also not part of the constituents A or B.
  • An additional constituent C can a peptide having more than one positively charged, ie basic, amino acid capable of traversing the membrane of a biological cell, wherein component C may, for example, comprise one or more different amino acids selected from the group consisting of arginine, lysine
  • a constituent C can also be understood, for example, to mean any other molecule which is different from the constituents A (active ingredient) and B (peptide) of the construct or is also no part of the components A or B and which is described in US Pat It is able to walk through the membrane of a biological cell.
  • the component C is a peptide having more than two amino acids selected from the group consisting of lysine, arginine and histidine, preferably more than three amino acids, preferably more than four amino acids, more preferably more than five amino acids, even more prefers more than six amino acids.
  • the peptide B connected to the active ingredient A is from 2 to 70 amino acids, preferably 2 to 50 amino acids, more preferably 2 to 30 amino acids and particularly preferably 2 to 25 amino acids are built up.
  • the construct according to the invention comprises several peptides B, these may be composed of the same number or a different number of amino acids.
  • Amino acids are organic acids which have at least one and usually not more than four amino groups NH 2 and at least one and usually not more than 4 carboxyl groups. Depending on the position of the amino group in the carbon chain to the terminal carboxyl COOH one distinguishes alpha-, beta-, gamma- amino acids, etc. (Encyclopedia of
  • amino acids is understood to mean all amino acids as defined above independently of the chirality that occurs. It should also be understood amino acids which have several chiral centers with the thus possible, different topological properties.
  • Basic amino acids have a side chain with a positive charge at pH 6, e.g. Arginine, lysine, histidine or other amino acids that possess these properties.
  • An acidic amino acid has a side chain with a negative charge at pH 6, such as glutamic acid, aspartic acid, phosphoserine, phosphothreonine or other amino acids that possess this property.
  • the peptide B of the active ingredient-peptide construct according to the invention can be composed of all amino acids which are known to the person skilled in the art as suitable for the purpose according to the invention and which bring about a drug-peptide construct having a net charge negative at pH 6.
  • the peptide B of the active ingredient-peptide construct according to the invention is composed of amino acids, which are selected from the group consisting of glutamic acid, aspartic acid, phosphoserine or phosphothreonine.
  • the construct according to the invention comprises several peptides B, these may be composed of the same or different amino acids.
  • the peptide B of the active ingredient-peptide construct according to the invention has an amino acid sequence selected from the group consisting of (Glu) 4 , (Glu) 5 , (Glu) 6 , (Glu) 7 , (Asp) 4 , ( Asp) 5 , (Asp) 6 , (Asp) 7 or sequences of lengths 4-7, which include Asp and Glu, regardless of the exact sequence of these amino acids.
  • amino acids which form peptides which are not or hardly degradable in the extracellular space, e.g. D-amino acids.
  • active ingredient A of the active ingredient-peptide construct according to the invention it is possible to use any active ingredient A which is known to the person skilled in the art as suitable for the purpose according to the invention.
  • active ingredients are suitable which are intended to exert their effect solely outside a biological cell and / or can. In the context of the invention, this includes on the one hand active substances which have a toxic effect or at least an undesired (side) effect within biological cells. On the other hand, however, they also include active substances which are ineffective inside a biological cell and whose concentration outside the cell has to be compensated for by migration into the cell by administration of increased doses.
  • active ingredients are suitable, which are better administered by linking with suitable peptides, for example, because not only their whereabouts in the extracellular space is ensured, but also their solubility can be improved.
  • the active ingredient A of the active ingredient-peptide construct according to the invention is a pharmaceutical active substance.
  • the active ingredient A is an active substance which has a positive net charge at pH 6.
  • Preferred active agents A of the active ingredient-peptide construct of the invention are effectors which can inhibit inflammatory processes in biological objects, preferably in animal and human medicine. Effectors of peptidyl-prolyl cis / trans isomerases (PPIases) are particularly preferred. In a further particularly preferred embodiment, the effectors comprise substances which inhibit the enzymatic activity of cyclophilin, and it is particularly preferred that, under inhibition, the reduction of the catalytic activity caused by the active ingredient under optimal conditions is understood to be at least 50%.
  • Effectors cause a variety of effects that can be used therapeutically. For example, they may have an influence on immunosuppression, neuroprotection / neurogeneration, chaperone activity, HIV infection, cancer or Alzheimer's disease.
  • PPIase inhibitors Although these effectors can be isolated between the individual PPIase families (Nature Chemical Biology, 3 (10): 619-29, 2007; Cellular & Molecular Life Sciences. 63 (24): 2889-900, 2006; Current Topics in Medicinal Chemistry. 3 (12): 1315-47, 2003; Advances in Protein Chemistry. 59: 243-82, 2001), but often have similar inhibitory power to sequence-like family members. Since PPIases within a family can influence a wide variety of biochemical reactions, the diagnostic or pharmacological effect of administered active ingredients directly depends on the concentration achieved in a wide variety of distribution areas. For example, some of these PPIase inhibitors (eg Biopolymers 84 (2006) 125-146; Chemical & Pharmaceutical Bulletin 54 (3): 372-376, 2006; Chemistry &
  • effectors include:
  • CSl peptides and its fragments which may affect the adherence of lymphocytes therapeutically desirable, as described in US 7,238,668.
  • inhibitors acting on TGF-beta such as NAALADase inhibitors, which regulate TGF-beta and on a variety of diseases such as neurodegenerative diseases, "Extracellular Matrix Formation Disorders", cell growth related diseases, infectious diseases, diseases of the immune system, "Epithelial Tissue Scarring ",” Collagen Vascular Diseases “,” Fibroproliferative Disorders ",” Connective Tissue Disorders ", inflammation, respiratory syndrome or infertility as shown for example in US 6,444,657, but also compounds which as described in US 6,693,118, on the extracellular TGF-beta - Concentration therapeutically usable act.
  • Cytokines such as Oncostatin-M or its biologically active fragments or protein constructs which have similar activity on tumor cells, e.g. in US 5,744,442 is summarized.
  • pyrrolo [3, 2-d] pyrimidine-2, 4-diones which act as adenosine receptor antagonists and can also act on inflammatory cytokines, as shown for example in US 7,449,473.
  • compounds which influence the action of secreted TNFalpha such as etanercept (Enbrel), inflixamab (Remicade), as shown, for example, in US Pat. No. 6,881,407.
  • Taorolidine and its medicinally active derivatives, which may affect the growth of tumors and metastases, e.g. in US Pat. No. 7,122,541.
  • PPIase inhibitors which are particularly preferably used as effectors (active ingredient A) in the context of the invention are:
  • Oligonucleotides or short nucleic acid molecules such as juglone or aromatic structures such as PIA, PIB PIC, PID, PIE, PIF, PIJ, as described in US 7,417,072 and Biopolymers 84 (2006) 125-146, respectively.
  • Pinl inhibitors can also therapeutically influence the disease-related expression of cytokines, as is observed, for example, in diseases of the asthmatic type or in the unwanted rejection of transplanted organs, or have an anticarcinogenic or antifungal action (eg: Cellular & Molecular Life Sciences 65 (2008) 359-375). Part of the Pinl effect may be due to the influence of TGFl-beta
  • FKBP binding pipecolic acid derivatives (US 6,500,843, US 6,022,878, US 5,846,981, US 5,843,960, US 5,801,197)
  • FKBP inhibitors are currently predominantly used or proposed to be used as agents with neurotrophic effects and are suitable for the treatment of nervous disorders (eg: WO 96/40140, WO 96/40633, PNAS 91 (1994) 3191-95, Nature Medicine 1 (1995 32-37, WO 96/40140, WO 96/40633, WO 97/16190, US 7276498) possibly caused by inhibition of FKBP-12 or FKBP-52, as an active substance with immunosuppressive properties for the prevention of transplant rejection (Clinical Chemistry 39 (Clin. Chem.
  • Tumor antigenic peptides and corresponding derivatives derived from cyclophilin B or cyclophilin (US 7,368,107, US 7,041,297).
  • Non-peptidic compounds suitable for therapeutically influencing the regeneration of neuronal cells (US Pat. No. 6,677,376).
  • HIV virus infections can affect therapeutically (US 6,593,362).
  • Non-immunosuppressive 6-position cyclosporin analogues (US 4,941,88).
  • 3-position cyclosporin derivatives particularly suitable for promoting hair growth (US 6,987,090, US 6,790,830, US 6,762,164).
  • Cyclosporin analogs particularly suitable for the treatment of autoimmune diseases (US 6,809,077).
  • Cyclosporin conjugates with amyloid-binding peptides for the treatment of neurological diseases such as Alzheimer's disease, multiple sclerosis or amyotrophic lateral sclerosis (US 6,316,405).
  • Cyclosporin derivatives particularly suitable for the treatment of HIV infections (US 5,948,884).
  • compositions for the treatment of transplant rejection, autoimmune or inflammatory diseases utilizing cyclosporin A and 40-O- (2-hydroxyethyl) rapamycin are provided.
  • Rapamycin derivatives (rapamycin and its derivatives for the treatment of neurological diseases and as a neuroprotective and neuro-regenerative substance (US Pat
  • Oxepane (US 5,344,833, US 5,221,740), imidazolyl derivatives (US 5,310,903), pyrazoles (US 5,169,851, US 5,164,399), Acetals (US 5,151,413), ethers (US 5,120,842), dimers (US 5,120,727) or hydrazones (US 5,120,726)
  • drug A of the drug-peptide construct of the present invention is selected from the group consisting of cyclosporin A, FK 506, and rapamycin.
  • the active ingredient A of the active ingredient-peptide construct according to the invention is an active substance which is sparingly soluble and in a particularly preferred embodiment the active ingredient A is an active substance which is sparingly soluble in the extracellular space.
  • “Poorly soluble active ingredient” is understood herein to mean a pharmaceutically active substance which has a solubility in water of less than 1 g (active ingredient) per 30 ml (water) at a temperature of 20 ° C.
  • the active ingredient A is cyclosporin A.
  • Cyclosporin (also cyclosporin) is a cyclic oligopeptide with immunosuppressive and calcineurin-inhibiting activity. It is characterized by a selective and reversible mechanism of immunosuppression. It selectively blocks the activation of T lymphocytes via the production of certain cytokines involved in the regulation of these T cells.
  • cyclosporin also cyclosporin is a cyclic oligopeptide with immunosuppressive and calcineurin-inhibiting activity. It is characterized by a selective and reversible mechanism of immunosuppression. It selectively blocks the activation of T lymphocytes via the production of certain cytokines involved in the regulation of these T cells.
  • Cyclosporin acts intracellularly by binding to the so-called cyclophilins or immunophilins belonging to the family of cyclosporin-binding proteins.
  • Inhibitors of cyclophilins have a very wide therapeutic spectrum, e.g. the treatment of diseases of the respiratory tract, e.g. Asthma, COPD, pneumonia or emphysema (Expert Opinion on Investigational Drugs 12 (2003) 647-653, Biodrugs 8 (1997) 205-215, American Journal of Respiratory Cell & Molecular Biology 20 (1999) 481-492), metabolic diseases such as diabetes
  • the complex of cyclosporin and cyclophilin subsequently blocks the serine-threonine phosphatase calcineurin. Their activity state in turn controls the activation of transcription factors such as NF-KappaB or NFATp / c, which play a crucial role in the activation of various cytokine genes, including interleukin-2.
  • transcription factors such as NF-KappaB or NFATp / c
  • interleukin-2 include interleukin-2.
  • T-heifer cells which increase the activity of the cytotoxic T cells responsible for the rejection, are the preferred target for cyclosporin.
  • cyclosporin inhibits the synthesis and release of other lymphokines responsible for the proliferation of mature cytotoxic T lymphocytes and other lymphocyte functions. Cyclosporin's ability to block interleukin-2 is clinical
  • the active ingredient A is FK 506 or rapamycin.
  • the peptide B is covalently bound to the active ingredient A.
  • the peptide can in principle be combined with the active compound A in any manner which is known to the person skilled in the art as suitable for the purpose according to the invention.
  • Peptide construct further include groups that provide the drug-peptide construct with other properties that are desirable for the skilled person for the particular application, as long as it is free of a component C as defined in more detail above.
  • the active ingredient-peptide construct according to the invention can be combined with one or more groups, which can be either the same, the same or different.
  • the inclusion of additional groups in the active ingredient -peptide construct according to the invention can serve on the one hand to already existing
  • the construct is also possible to provide the construct with new, additional properties. It is conceivable, for example, that the construct is provided with an indicator in order to control its accumulation in the desired tissue or to be able to classify the desired tissue by means of indicator distribution. Furthermore, it is conceivable that the construct is provided with a group that allows its accumulation in very specific tissues.
  • the drug-peptide construct comprises an indicator.
  • the indicator is covalently bonded to the active ingredient A.
  • the indicator can in principle be combined with the active substance A in any manner which is known to the person skilled in the art as suitable for the purpose according to the invention.
  • the indicator is covalently bound to the peptide B.
  • the indicator may in principle be linked to the peptide B in any manner known to those skilled in the art as being suitable for the purpose of the invention.
  • the indicator is covalently bound to a linker which links peptide B to drug A.
  • the linker may be any compound which is known to the person skilled in the art as suitable for the purpose according to the invention. However, it is preferably a compound which is free from a protease cleavage site. Particularly preferably, the linker is selected from the group consisting of molecules which form an atomic distance between four and 40 atoms.
  • the term "indicator” means substances such as, preferably, dyes, voltage-sensitive indicators, pH indicators, calcium-sensitive indicators, radioactive elements, NMR labels or electron spin labels, which have been described several times in the scientific literature ( WO / 2005/022158, EP 0649022, US Pat. No. 6,596,499, US Pat. No. 7,090,995, US Pat. No. 4,672,044)
  • the term indicator preferably comprises individual atoms or molecules which are covalently bonded to the Construct are connected.
  • indicator or else several indicators can be covalently bound directly to the active ingredient molecule, but the indicator or else several indicators can also be bound to a multifunctional linker or the indicator or else several indicators can also be present within the acidic peptide or terminally on the acidic peptide be covalently bound.
  • indicator preferably includes dyes, stress-sensitive indicators, pH-sensitive indicators, radioactive elements, calcium-sensitive indicators, NMR labels and electrospins labels.
  • Dyes in the context of the invention are substances that can be detected optically by detecting the emitted by them or not absorbed by them electromagnetic radiation. These include e.g. Dyes such as fluorescein isocyanate (FIC), fluorescein isothiocyanate (FITC), dimethylaminonaphthalene-S-sulphonyl chloride (DANSC), tetramethylrhodamine isothiocyanate (TRITC), lissamine rhodamine B200 sulphonylchloride (RB 200 SC), etc.
  • FIC fluorescein isocyanate
  • FITC fluorescein isothiocyanate
  • DANSC dimethylaminonaphthalene-S-sulphonyl chloride
  • TRITC tetramethylrhodamine isothiocyanate
  • RB 200 SC lissamine rhodamine B200 sulphonylchloride
  • Voltage-sensitive indicators in the sense of the invention are substances which, depending on an applied electrical potential difference or the present electrical potential, change their physical, optical or catalytic properties in such a way that they produce a detectable signal.
  • Those skilled in the art are known voltage-sensitive indicators such.
  • pH-sensitive indicators in the sense of the invention are substances which, depending on the pH, change their physical, optical or catalytic properties in such a way that they produce a detectable signal.
  • indicator dyes such as phenol red
  • Bromothymol blue, bromophenol blue u.v.a. are known in the art.
  • Calcium-sensitive indicators in the context of the invention are substances which, in the presence of calcium, change their physical, optical or catalytic properties such that they produce a detectable signal.
  • the calcium-sensitive indicators known to the expert are z.
  • Radioactive elements in the context of the invention produce eg gamma radiation, such as the following isotopes 124 J, 125 J, 128 J, 131 J, 1 32 J or 51 Cr, with particular preference being given to the 125 J.
  • Others such as 11 C, 18 F, 15 O or 13 N, can by means of their
  • Positron radiation and corresponding detectors positron emission tomography
  • others such as 111 In
  • NMR labels in the sense of the invention are substances in which atoms with an odd number of nucleons (sum of the protons and neutrons) are contained Such atomic nuclei, for example: 13 C, 15 N or 19 F possess a nuclear spin and thus a nuclear magnetic moment.
  • Electron spin label serve in the context of the invention, the measurement of "Electron Paramagnetic Resonance” by means of electron spin resonance. This is the resonant Microwave absorption of a sample measured in an external magnetic field. This makes it possible to detect molecules which have a permanent magnetic moment (unpaired electrons) (Physics in Medicine & Biology, 43 (1998) U 3-U 4, Clinical Chemistry & Laboratory Medicine, 46 (2008) 1203-1210).
  • the active ingredient-peptide construct according to the invention may contain one or more indicators which may be of the same or different nature.
  • indicators is particularly advantageous when the active ingredient -peptide construct of the invention is to be used for the manufacture of a medicament for use in a therapeutic method such as a diagnostic method (e.g., history taking, physical examination, use of imaging techniques such as X-ray / MRI or Analysis with laboratory values of the blood and other body fluids).
  • a diagnostic method e.g., history taking, physical examination, use of imaging techniques such as X-ray / MRI or Analysis with laboratory values of the blood and other body fluids.
  • the active ingredient-peptide constructs according to the invention contain one or more indicators, the distribution space of the active substance A can be recognized on the basis of these indicators.
  • indicators can be used to quantify drug A.
  • the active ingredient-peptide construct according to the invention is furthermore free of a protease cleavage site, in particular free of one
  • a protease cleavage site which, after cleavage, cleaves peptide B from the construct, such as a linker of appropriate amino acids (as known to those of skill in the art), which could serve as the compound of the individual groups of the construct, for example.
  • the active ingredient-peptide construct according to the invention relates, in particularly preferred embodiments, to constructs or a molecule having the following formulas, in which R, if present, is a carboxy-Tamra or acetyl radical.
  • the invention relates to a method for the accumulation of active substances in an extracellular space of a multicellular object, comprising the steps:
  • extracellular space is to be understood as meaning all the areas which are located outside the cytosol and the membrane enclosing the cytosol, including, for example, the culture solution present in cell suspensions.
  • the multicellular object can be any object that consists of at least two identical or different biological cells.
  • biological cell encompasses both human, animal, plant and bacterial cells as well as unicellular organisms.
  • the term "multicellular object” means an accumulation of several cells, such as a cell colony of a bacterial culture understood.
  • multicellular object means a separated body part such as a graft, especially an organ, a body part such as a limb or a tissue graft, blood or a blood fraction such as for example, blood plasma or an in vitro culture of human and / or animal cells, such as a two-dimensional tissue culture or a spheroid culture understood the cells.
  • the term “multicellular object” is understood as meaning a part of a plant, such as, for example, leaves, roots or stems, or even an entire plant.
  • the multicellular object is a separated organ or body part, blood or a blood fraction, a cell culture or a plant.
  • the invention further relates to the use of the active ingredient-peptide construct according to the invention as a medicament.
  • the construct according to the invention can be used for the production of medicaments.
  • the construct according to the invention is preferably used for the treatment of non-immunosuppressive diseases.
  • Field of application of the medicaments according to invention can be therapy and diagnosis of illnesses but also of cosmetic kind, whereby under therapy in the broadest sense also the
  • Control of pests in the animal and plant kingdom or the support of healing processes in the animal and plant kingdom but also the influence of biological processes should be understood in the desired manner.
  • Particular advantages are in animal and human medicine, in the application of substances on or in cell suspensions, tissue cultures, transplants or the entire mammals.
  • the present invention further relates to the use of the active ingredient-peptide construct according to the invention as a diagnostic agent.
  • the invention further relates to the use of the construct according to the invention for the manufacture of a medicament for the treatment of non-immunosuppressive diseases.
  • the medicament may be administered in any form known to the person skilled in the art as suitable for the intended purpose.
  • the drug may be used in a form selected from the group consisting of injections, infusions, tablets, creams, sprays, capsules, syrups, emulsions, powders, powders, suppositories, or the like. It is particularly preferred that the drug is used in the form of sprays or tablets.
  • the present invention further relates in a further aspect to a pharmaceutical composition comprising an active ingredient-peptide construct according to the invention.
  • the composition may be any pharmaceutical composition known to those skilled in the art.
  • the pharmaceutical composition is sprays or tablets.
  • Fig. 2 The cyclosporin derivative Cs9-TAMRA
  • Fig. 3 The cyclosporin derivative CsMl
  • Fig. 4 The cyclosporin derivative CsM2
  • Fig. 5 The cyclosporin derivative CsM3
  • FIG. 6 The trifunctional linker (MM-50)
  • FIG. 7 The trifunctional linker (MM-50) linked to TAMRA
  • Fig. 8 The cyclosporin derivative MM-218
  • Fig. 10 The cyclosporin derivative CsM4
  • FIG. 11 The cyclosporin derivative CsM5
  • FIG. 12 The cyclosporin derivative CsM6, wherein R stands for a carboxy-TAMRA or an acetyl radical.
  • Fig. 13 The cyclosporin derivative CsM7
  • Fig. 15 The FK506 derivative FKM2
  • FIG. 16 The FK506 derivative FKM3, wherein R stands for a carboxy-TAMRA or acetyl radical.
  • Fig. 17 The FK506 derivative FKM4
  • Fig. 18 The rapamycin derivative RPMl
  • Fig. 19 The rapamycin derivative RPM2
  • Fig. 20 The rapamycin derivative RPM3, wherein R is a
  • Carboxy-TAMRA or acetyl radical Carboxy-TAMRA or acetyl radical.
  • Fig. 21A, B Control images: Heia cells without added
  • Cyclosporin derivative recorded by means of phase contrast (A) and fluorescence (B).
  • Fig. 21C, D MM218 incubation: Heia cells incubated with 250 nM MM218 for 2 h recorded by phase contrast (C) and fluorescence (D).
  • Fig. 2IE, F Cs9-Rhh incubation: Heia cells incubated with 250 nM Cs9-Rhd taken for 2 h by means of phase contrast
  • FIG. 22 Influence of MM218 on the number of CD4-positive T cells which migrated into the bronchial mucous membranes through ovalbumin sensitization.
  • Fig. 23 Influence of MM218 on the number of eosinophils granulocytes (eosinophils), which are caused by the
  • Fig. 24 Chemotaxis assay. It is shown that without any addition of a stimulus (-) a chemotaxis index of about 2.7 +/- 0.3 is obtained.
  • MALDI mass spectrometry gave a mass [MH-H] + of 1461.3 (calculated: 1460). Subsequently, the substance was incubated with 5 ml of ZnCl 2 / ether under nitrogen contactor for three hours at room temperature. After adding 0.1 ml of water and 15 ml of acetonitrile, the precipitated salt was filtered off. After vacuum drying and separation by C8 HPLC, a product of mass [M + H] + of 1361.3 (theoretical: 1360.1) could be obtained by MALDI mass spectrometry. Subsequently, 40 mg of this substance (Cs9) was mixed for 15 minutes
  • Cyclosporin derivative CsMl ( Figure 3) CsMl was prepared on a 2-ClTrt matrix using standard Fmoc procedures. In each cycle, the Fmoc-protected amino acids are activated with PyBOP and DIPEA in DMF and then coupled for two hours. The Fmoc protecting group is split off in each case with 20% piperidine in DMF. The Tamra-labeled trifunctional linker was coupled overnight as described above. The cyclosporin derivative (Cs6) was pre-activated and coupled with HATU, HOAt and DIPEA and overnight. The side chain of D-glutamic acid was protected as a t-butyl ester. After synthesis, the product was removed from the matrix by 50%.
  • CsM2a 100 mg of Cs6, 20 parts of NH 2 (CH 2 -CH 2 -O) 2 CH 2 CH 2 NH 2 and 1.1 parts of PyBop were stirred in 5 ml of DMF at room temperature overnight. Subsequently, 40 ml of ethyl acetate were added and the organic layer washed with 5% NaHSO 4 , 5% NaHCO 3 and saturated NaCl solution. After drying with Na 2 SO 4 and subsequent vacuum drying, the product (CsM2a) was separated by HPLC. Subsequently, CsM2a was stirred with 5 parts of succinic anhydride and 10 parts of DIPEA in 5 ml of DMF at room temperature overnight.
  • MM-218 was prepared on a 2-ClTrt matrix using standard Fmoc procedures.
  • the Fmoc-protected amino acids are activated with PyBOP and DIPEA in DMF and then coupled for two hours.
  • the Fmoc protecting group is split off in each case with 20% piperidine in DMF.
  • the Tamra-labeled trifunctional linker was coupled overnight as described above.
  • the cyclosporin derivative (Cs6) was pre-activated and coupled with HATU, HOAt and DIPEA and overnight.
  • the side chain of D-glutamic acid was protected as a t-butyl ester.
  • H-Dap fluorescein
  • D-Glu D-Glu 6 -Gly-OH
  • Fmoc-protected amino acids were first pre-activated with PyBOP and DIPEA in DMF and subsequently coupled for two hours.
  • the side chain of D-glutamic acid was protected as a t-butyl ester.
  • the Fmoc protecting group was cleaved by means of 20% piperidine in DMF.
  • the product was cleaved from the matrix with 50% TFA / CH 2 Cl 2 at room temperature and isolated by RP HPLC.
  • MALDI mass spectrometry a mass [M + H] + of 1294.3 (calculated 1293.4) could be determined.
  • H-Dap (fluorescein) - (D-Glu) 6 -Gly-OH was added to a solution of Cyclosporin derivative-6 (Cs6) in DMF to which 0.9 parts of HATU and 3 parts of DIPEA were added and mixed for 30 minutes , added and stirred overnight.
  • Cs6 Cyclosporin derivative-6
  • cyclosporin at position 1 Derivatization of the cyclosporin at position 1 is achieved by boiling cyclosporin A and 0.1 part of the "Hoveyda Grubbs catalyst second generation" (1,3-bis- (2,4,6-trimethylphenyl) -2-imidazolidinylidenes). chloro (o-isopropoxyphenylmethylene) ruthenium) and 20 parts of dimethyl maleate in toluene under reflux for 45 hours. Then the toluene is removed in vacuo and the residue is dissolved in DCM / MeOH (10: 0.5) and filtered through silica gel.
  • Hoveyda Grubbs catalyst second generation (1,3-bis- (2,4,6-trimethylphenyl) -2-imidazolidinylidenes).
  • the peptide is synthesized on a 2-ClTrt matrix using conventional Fmoc chemistry.
  • Fmoc-protected amino acids are coupled with PyBOP and DIPEA in DMF for two hours.
  • the Fmoc protecting group is split off in each case with 20% piperidine in DMF.
  • the trifunctional linker (Example Ic) is coupled overnight.
  • the side chain of D-glutamic acid is protected as t-butyl ester.
  • Cs6 ( Figure 1) is added with HATU, HOAt and DIPEA and stirred overnight.
  • the product CsM6 can be obtained after cleavage from the matrix with 50% TFA / CH 2 Cl 2 at 5 0 C and purification by RP HPLC.
  • CsM4 ( Figure 10), 20 parts of NH 2 (CH 2 -CH 2 -O) 2 CH 2 CH 2 NH 2 and 1.1 parts of PyBop in DMF are stirred at room temperature overnight. Then 40 ml of ethyl acetate are added and the organic layer is washed with 5% NaHSO 4 , 5% NaHCO 3 and saturated NaCl solution. After drying with Na 2 SO 4 and subsequent vacuum drying, the product (CsM7a) is separated by means of HPLC. Subsequently, CsM7a is stirred with 1 part HATU, 3 parts DIPEA in 3 ml DMF for 10 min, at room temperature. Thereafter, the solution is added to a mixture of an equivalent part of H (D-Glu) 6- Gly-OH dissolved in 2 ml of DMF and stirred overnight. To
  • FKM1 (FIG. 14) 1 part of HATU and 3 parts of DIPEA are mixed with 3 ml of DMF and stirred for 20 minutes at room temperature. Subsequently, a solution containing one part of H- (D-Glu) 6- Gly-OH in 2 ml of DMF is added and stirred overnight. After filtering off insoluble constituents, the product (FKM2) can be obtained by means of preparative HPLC.
  • the peptide is synthesized on a 2-CITrt matrix using conventional Fmoc chemistry.
  • Fmoc-protected amino acids are first pre-activated with PyBOP and DIPEA in DMF and subsequently coupled for 2 h.
  • the trifunctional linker (Example Ic) is coupled overnight.
  • the Fmoc protecting group is split off in each case with 20% piperidine in DMF.
  • the side chain of D-glutamic acid is protected as a t-butyl ester.
  • FKMl (Fig. 14) is added to HATU, HOAt and DIPEA and stirred overnight.
  • the product FKM3 can be obtained after cleavage from the matrix with 50% TFA / CH 2 Cl 2 at 5 0 C and purification by RP HPLC.
  • FKMl ( Figure 14), 20 parts of NH 2 (CH 2 -CH 2 -O) 2 CH 2 CH 2 NH 2 and 1.1 parts of PyBop in DMF are stirred at room temperature overnight. Then 40 ml of ethyl acetate are added and the organic layer is washed with 5% NaHSO 4 , 5% NaHCO 3 and saturated NaCl solution. After drying with Na 2 SO 4 and subsequent vacuum drying, the product (FKM4a) is separated by means of HPLC. Subsequently, FKM4a are stirred with 5 parts of succinic anhydride and 10 parts of DIPEA in 5 ml of DMF overnight at room temperature.
  • the product FKM4 can be obtained.
  • Rapamycin derivative RPMl Figure 18
  • a solution of one part of rapamycin, 5 parts of 2,6-lutidine and 5 parts of bromoethyl triflate are incubated in toluene at 65 0 C for 18 h. After cooling, saturated sodium bicarbonate solution is added and the product extracted with ethyl acetate. The extraction is repeated three times. The combined extracts are filtered and dried in vacuo. Subsequently, the product (RPMIa) is isolated by means of preparative HPLC and taken up in DMF. After adding 1.2 parts of sodium azide, the mixture is stirred for two hours. After adding saturated sodium bicarbonate solution, the solution is washed three times
  • RPMIb Acetylacetate extracted. The combined extracts are then dried over Na 2 SO 4 , filtered and dried by vacuum.
  • the product (RPMIb) can then be obtained by means of preparative HPLC. Thereafter, RPMIb is taken up in 70% THF and stirred after addition of five parts of triphenylphosphine overnight. After addition of ethyl acetate, the solution is washed three times with saturated brine and then dried over Na 2 SO 4 and filtered.
  • the product (RPMIc) is then isolated by pre-pertinent HPLC. After dissolving the RPMIc in DMF, 1.1 parts of succinic anhydride are added and the pH of the solution is adjusted to about pH 7.5 with diisopropylethylamine. After stirring the mixture overnight, the product RPM1 can be obtained by means of preparative HPLC.
  • RPM1 one part of HATU and 3 parts of DIPEA are mixed with 3 ml of DMF and stirred for 20 min at room temperature. Subsequently, a portion of H- (D-Glu) 6- Gly-OH in 2 ml of DMF is added and stirred overnight. After filtering off undissolved residues, the product RPM2 can be isolated by means of preparative HPLC. c) rapamycin derivative RPM2 ( Figure 20)
  • the compound is synthesized on a 2-CITrt matrix using conventional Fmoc chemistry.
  • the Fmoc-protected amino acids are activated with PyBOP and DIPEA in DMF with a coupling time of two hours.
  • the Fmoc protecting group is split off each with 20% piperidine in DMF.
  • the trifunctional linker is coupled overnight.
  • the rapamycin derivative RPM1 is coupled overnight using HATU, HOAt and DIPEA.
  • Glutamic acid side chain is protected as t-butyl ester. Subsequently, the peptide is cleaved from the matrix by means of 50% TFA / CH 2 Cl 2 at 5 0 C and isolated by RP HPLC.
  • the advantage of the present invention becomes clear in the study of the transport behavior of two cyclosporin derivatives.
  • the derivative Cs9 derivative MM218 differs from the Cs9 derivative Cs9-Rhd by the synthesized acidic peptide.
  • the uptake of chemically modified fluorescent CsA derivatives into living eukaryotic cells is demonstrated on a cell line by confocal laser scanning microscopy.
  • 10 5 Heia cells were used in Petri dishes from Ibidi® ( ⁇ -Dish, 35 mm, high) and incubated for 1-2 days in DME medium (high glucose) at 37 ° C. and 5% CO 2 .
  • the examination was carried out on an inverted microscope (Nikon ECLIPSE C1TE2000-E) equipped with a focusing aid, the T-PFS, to prevent a so-called focus drift.
  • the images were taken with a phase contrast objective (40,0x Plan Fluor Oilimmersion NA 1,30) used as well as the microscope own software EZ-Cl 3.7.
  • the fluorophore 5- (6) -carboxytetramethylrhodamine was excited by a 561 nm laser from Melles & Griot.
  • the cells were first washed twice with 2 ml PBS pH 7.4 (Dulbecco) and then taken up in 2 ml MIK medium (phenol red free DME medium, FCS-free, with 20 mM HEPES pH 7.2 and 0.01 % Carbencilin) and incubated for 20 min at 37 0 C and 5% CO 2 .
  • MIK medium phenol red free DME medium, FCS-free, with 20 mM HEPES pH 7.2 and 0.01 % Carbencilin
  • Fig. 20 shows Heia cells incubated with CsA derivatives after 2 h.
  • Fig. 2OA, B control images: Heia cells without added Cyclosporinderivat by means of phase contrast (A) and fluorescence (B). In fluorescent light no structures are visible.
  • Fig. 2OC, D MM218 Incubation: Heia cells incubated with 250 nM MM218 for 2 h, using phase contrast (C) and fluorescence (D). In fluorescent light, the Heia cells are visible only as shadows in the vicinity of the fluorescent cyclosporin derivative. The acidic peptide cyclosporin derivative is not transported to Heia cells.
  • Fig. 2OA, B control images: Heia cells without added Cyclosporinderivat by means of phase contrast (A) and fluorescence (B). In fluorescent light no structures are visible.
  • Fig. 2OC, D MM218 Incubation: Heia cells incubated with 250 nM MM218 for 2
  • Cs9-Rhd incubation Heia cells incubated with 250 nM Cs9-Rhd for 2 h by means of phase contrast (E) and fluorescence (F). In fluorescent light, the Heia cells are visible as fluorescent cells. The non-acidic peptide cyclosporin derivative accumulates within the heia cells.
  • mice Removed spleen from mice (BALB / c line) was squeezed between slides to prepare appropriate cell suspensions. Subsequently, the suspension thus obtained was filtered through a nylon screen to separate coarse matter. The cells thus obtained were centrifuged together with a lymphocyte separation medium (Mediatech) to obtain mononuclear cells.
  • a lymphocyte separation medium Mediatech
  • mice Female mice (BALB / c line) were sensitized by intraperitoneal (i.p.) administration of 50 ⁇ g ovalbumin in phosphate buffer (PBS) (OVA) plus 100 ⁇ L aluminum hydroxide (alum) at a total volume of 200 ⁇ L per mouse on day 0. The OVA / alum-sensitized mice were then treated under mild anesthesia
  • FIG. 23 Influence of MM218 on the number of eosinophilic granulocytes (eosinophils), which migrated into the bronchial mucous membranes through ovalbumin sensitization.
  • the untreated mice (naive) also served as controls, as did the OVA-sensitized (-) and MM218 solvent-only treated animals.
  • MM218 significantly reduced the number of eosinophils.
  • Activated CD4 + T cells were obtained by stimulation of the mononuclear cells (3 ⁇ 10 6 cells / well plate cavity) generated in the proliferation assay with ConA (10 ⁇ g / ml) overnight. Subsequently, the CD4 + T cells were analyzed by MACS negative selection kit (Miltenyi Biotec, Auburn Ca). The chemotaxis assay was performed in Boyden chambers (Neuroprobe) with 48 wells, each well consisting of two compartments separated by a 5- ⁇ m polycarbonate membrane (Neuroprobe). The assays were started by adding 10 4 cells to the upper compartment medium (RPMI 1640 + 1% BSA). The lower compartment medium contained either 100 ng / ml human cyclophilin A (Calbiochem) or no additives. The effect of drugs on cell migration was compared after addition of these compounds to both compartments. The used
  • Drug concentration was either 2 ⁇ M MM218 dissolved in ethanol or 1% ethanol (Diluent). Subsequently, the thus-loaded chemotaxis chambers were incubated at 37 ° C. in 5% CO 2 for 50 minutes. Thereafter, the separation membrane was removed and cells that had not migrated were scraped off. Subsequently, the membrane was stained with Wright-Giemsa solution (CAMCO, Fort Lauderdale, FL). The chemotactic index was then obtained for each membrane by dividing the number of cells migrated by the number of cells migrating without any stimulation. Figure 24 shows that without any addition of stimulus (-), a chemotaxis index of about 2.7 +/- 0.3 is obtained. The addition of the MM218 solvent (+ Diluent) shows a non-significant, the addition of the drug has a highly significant influence on the chemotaxis index.

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Abstract

La présente invention concerne une construction à base d'un principe actif et d'un peptide pour une administration extracellulaire, un procédé pour administrer des principes actifs dans un espace extracellulaire d'un objet multicellulaire, l'utilisation de la construction de principe actif et de peptide selon l'invention pour fabriquer un médicament, ainsi qu'une composition pharmaceutique contenant la construction de principe actif et de peptide selon l'invention.
EP09774835A 2008-12-04 2009-12-04 Construction de principe actif et de peptide pour une administration extracellulaire Withdrawn EP2370106A2 (fr)

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DE102011111991A1 (de) 2011-08-30 2013-02-28 Lead Discovery Center Gmbh Neue Cyclosporin-Derivate
EP2705856A1 (fr) * 2012-09-07 2014-03-12 Deutsches Zentrum für Neurodegenerative Erkrankungen e.V. Composés pour le traitement de troubles neurodégénératifs

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1999030164A1 (fr) * 1997-12-09 1999-06-17 President And Fellows Of Harvard College Procede d'identification de modulateurs de transcription
US6030941A (en) * 1996-05-01 2000-02-29 Avi Biopharma, Inc. Polymer composition for delivering substances in living organisms

Family Cites Families (91)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US598325A (en) 1898-02-01 Gomeey mcintyre
US494188A (en) 1893-03-28 Allen g
US723286A (en) 1901-12-02 1903-03-24 James J Lawler Check-valve for hot-water systems.
US985890A (en) 1910-07-25 1911-03-07 Jonathan O Fowler Weighing and vending machine.
SE448386B (sv) 1978-10-18 1987-02-16 Sandoz Ag Nya cyklosporinderivat, forfarande for framstellning av dem samt farmaceutisk komposition innehallande dem
US4261989A (en) 1979-02-19 1981-04-14 Kaken Chemical Co. Ltd. Geldanamycin derivatives and antitumor drug
US4673678A (en) 1986-07-25 1987-06-16 The United States Of America As Represented By The Secretary Of The Department Of Health And Human Services Water soluble derivatives of fredericamycin A
US4885276A (en) 1987-06-03 1989-12-05 Merck & Co., Inc. Cyclosporin analogs with modified "C-9 amino acids"
US4798823A (en) 1987-06-03 1989-01-17 Merck & Co., Inc. New cyclosporin analogs with modified "C-9 amino acids"
DE3851268T2 (de) 1987-06-19 1995-01-26 Sandoz Ag Zyklische Peptolide.
EP0373260B1 (fr) 1987-06-22 1994-03-09 Merck & Co. Inc. Dérivés de cyclosporine avec un acide aminé modifié en position 8
US4914188A (en) 1987-11-16 1990-04-03 Merck & Co., Inc. Novel 6-position cyclosporin analogs as non-immunosuppressive antagonists of cyclosporin binding to cyclophilin
DK169306B1 (da) 1989-06-14 1994-10-10 Af 18 Juni 1990 As Middel til fjernelse eller inaktivering af uønskede komponenter i blod eller andre ekstracellulære legemsvæsker samt fremgangsmåde til fremstilling af dextran-O-carbonyl-benzo-18-crown-6 og af dextran-hydroxyethylbenzo-18-crown-6
US7238668B1 (en) 1989-09-01 2007-07-03 Fred Hutchinson Cancer Research Center Inhibition of lymphocyte adherence with CS-1-peptides and fragments thereof
US5120649A (en) 1990-05-15 1992-06-09 New York Blood Center, Inc. Photodynamic inactivation of viruses in blood cell-containing compositions
US5733892A (en) 1990-07-24 1998-03-31 Seikagaku Corporation Metastasis inhibitor composition comprising a phospholipid-linked glycosaminoglycan and method for inhibiting metastasis employing the same
US5166208A (en) 1991-10-09 1992-11-24 Boston College Fredericamycin A derivatives
US5744442A (en) 1992-08-26 1998-04-28 Bristol Meyers Squibb Company Regulation of cellular invasiveness
US6770279B1 (en) 1992-10-08 2004-08-03 The Kennedy Institute Of Rheumatology TNFα antagonists and cyclosporin in therapy of rheumatoid arthritis
US5846981A (en) 1993-05-28 1998-12-08 Gpi Nil Holdings Inc. Inhibitors of rotamase enzyme activity
IL112873A (en) 1994-03-08 2005-03-20 Wyeth Corp Rapamycin-fkbp12 binding proteins, their isolation and their use
DE4430127A1 (de) 1994-08-25 1996-03-14 Hoechst Ag Kombinationspräparat, enthaltend Cyclosporin A oder FK506 und ein Xanthinderivat
US5696135A (en) 1995-06-07 1997-12-09 Gpi Nil Holdings, Inc. Inhibitors of rotamase enzyme activity effective at stimulating neuronal growth
US5859031A (en) 1995-06-07 1999-01-12 Gpi Nil Holdings, Inc. Small molecule inhibitors of rotamase enzyme activity
US6291510B1 (en) 1995-06-07 2001-09-18 Gpi Nil Holdings, Inc. Small molecule inhibitors of rotamase enzyme activity
US6509477B1 (en) 1998-11-12 2003-01-21 Guilford Pharmaceuticals Inc. Small molecule inhibitors of rotamase enzyme activity
KR100400620B1 (ko) 1995-06-09 2004-02-18 노파르티스 아게 라파마이신유도체
EP0842191B1 (fr) 1995-07-17 2001-11-21 C-Chem AG Derives de cyclosporin ayant une activite contre hiv
US5801197A (en) 1995-10-31 1998-09-01 Gpi Nil Holdings, Inc. Rotamase enzyme activity inhibitors
NZ333657A (en) 1996-07-30 2000-05-26 Novartis Ag A pharmaceutical composition comprising cyclosporin A and 40-O-(2-hydroxyethyl)-rapamycin
US6001965A (en) 1996-11-21 1999-12-14 The Regents Of The University Of Michigan Anticancer compounds and methods
FR2757522B1 (fr) 1996-12-24 1999-01-29 Rhone Poulenc Rorer Sa Derives de cyclosporine, leur preparation et les compositions pharmaceutiques qui les contiennent
FR2757521B1 (fr) 1996-12-24 1999-01-29 Rhone Poulenc Rorer Sa Nouveaux derives de cyclosporine, leur preparation et les compositions pharmaceutiques qui les contiennent
US5989591A (en) 1997-03-14 1999-11-23 American Home Products Corporation Rapamycin formulations for oral administration
US6709873B1 (en) 1997-04-09 2004-03-23 Isodiagnostika Inc. Method for production of antibodies to specific sites of rapamycin
US6273913B1 (en) 1997-04-18 2001-08-14 Cordis Corporation Modified stent useful for delivery of drugs along stent strut
FR2762843B1 (fr) 1997-04-30 1999-12-10 Rhone Poulenc Rorer Sa Nouveaux derives de cyclosporine, leur preparation et les compositions pharmaceutiques qui les contiennent
US6916790B2 (en) 1997-05-05 2005-07-12 Mayo Foundation For Medical Education And Research Treatment of osteoporosis
FR2763847B1 (fr) 1997-05-28 2003-06-06 Sanofi Sa Utilisation de tetrahydropyridines 4-substituees pour fabriquer des medicaments agissant sur le tgf-beta-1
WO1999010374A1 (fr) 1997-08-26 1999-03-04 Wisconsin Alumni Research Foundation Conjugues de cyclosporine a et utilisations
US6890546B2 (en) 1998-09-24 2005-05-10 Abbott Laboratories Medical devices containing rapamycin analogs
US20030220234A1 (en) 1998-11-02 2003-11-27 Selvaraj Naicker Deuterated cyclosporine analogs and their use as immunodulating agents
AU749544B2 (en) 1998-06-25 2002-06-27 Dainippon Sumitomo Pharma Co., Ltd. Tumor antigen peptides originating in cyclophilin B
USRE40987E1 (en) 1998-07-01 2009-11-17 Debiopharm S.A. Cyclosporin with improved activity profile
GB9815696D0 (en) 1998-07-20 1998-09-16 Pfizer Ltd Heterocyclics
GB9815880D0 (en) 1998-07-21 1998-09-16 Pfizer Ltd Heterocycles
US7410995B1 (en) 1998-08-14 2008-08-12 Gpi Nil Holdings Inc. N-linked sulfonamide of heterocyclic thioesters for vision and memory disorders
US7338976B1 (en) 1998-08-14 2008-03-04 Gpi Nil Holdings, Inc. Heterocyclic esters or amides for vision and memory disorders
US7265150B1 (en) 1998-08-14 2007-09-04 Gpi Nil Holdings Inc. Carboxylic acids and carboxylic acid isosteres of N-heterocyclic compounds for vision and memory disorders
US6015809A (en) 1998-08-17 2000-01-18 American Home Products Corporation Photocyclized rapamycin
DE19859910C2 (de) 1998-12-23 2001-03-22 Ratiopharm Gmbh Orales Arzneimittel
US6444657B1 (en) 1998-12-31 2002-09-03 Guilford Pharmaceuticals Inc. Methods for treating certain diseases using naaladase inhibitors
US20010041189A1 (en) * 1999-04-13 2001-11-15 Jingya Xu Poly(dipeptide) as a drug carrier
US6495549B1 (en) 1999-07-15 2002-12-17 Pfizer Inc FKBP inhibitors
US6730293B1 (en) * 1999-08-24 2004-05-04 Cellgate, Inc. Compositions and methods for treating inflammatory diseases of the skin
AU7353300A (en) 1999-09-08 2001-04-10 Guilford Pharmaceuticals Inc. Non-peptidic cyclophilin binding compounds and their use
US6277983B1 (en) 2000-09-27 2001-08-21 American Home Products Corporation Regioselective synthesis of rapamycin derivatives
EP1466627A1 (fr) * 1999-10-12 2004-10-13 Cell Therapeutics, Inc. Polyglutamate-camptothecin conjugés
US20020014242A1 (en) 2000-07-31 2002-02-07 Abraham Scaria Use of rapamycin to inhibit immune response and induce tolerance to gene therapy vector and encoded transgene products
US6881407B2 (en) 2000-08-11 2005-04-19 Ashok Amin Method for treating hepatitis
US7163918B2 (en) * 2000-08-22 2007-01-16 New River Pharmaceuticals Inc. Iodothyronine compositions
US6656971B2 (en) 2001-01-25 2003-12-02 Guilford Pharmaceuticals Inc. Trisubstituted carbocyclic cyclophilin binding compounds and their use
PE20020908A1 (es) * 2001-03-21 2002-10-26 Cell Therapeutics Inc Produccion recombinante de polimeros polianionicos y uso de de los mismos
MXPA03007856A (es) 2001-03-30 2003-12-04 Us Health Derivado de geldanamicina y metodo de usarlo para tratar cancer.
KR100465012B1 (ko) 2001-05-11 2005-01-13 주식회사 엘지생활건강 모발 성장 효과를 갖는 사이클로스포린 3 위치 유도체를유효성분으로 하는 모발 성장 촉진제
US6593362B2 (en) 2001-05-21 2003-07-15 Guilford Pharmaceuticals Inc. Non-peptidic cyclophilin binding compounds and their use
ITMI20011057A1 (it) 2001-05-22 2002-11-22 Bracco Imaging Spa Preparazione ed uso di peptidi ciclici e ramificati e loro deriati marcati come agenti terapeutici agonisti o antagonisti della colecistochi
US7034037B2 (en) 2001-06-29 2006-04-25 Ethicon, Inc. Compositions and medical devices utilizing bioabsorbable polymeric waxes and rapamycin
US6566357B1 (en) 2001-09-20 2003-05-20 The Procter & Gamble Co. Spirocyclic-6,7-dihydro-5H-pyrazolo[1,2-a]pyrazol-1-ones which control inflammatory cytokines
US6730668B2 (en) 2001-09-20 2004-05-04 The Procter & Gamble Company 6,7-dihydro-5H-pyrazolo[1,2-a]pyrazol-1-ones which control inflammatory cytokines
JP5280603B2 (ja) 2001-10-01 2013-09-04 ガイストリッヒ ファーマ アーゲー 転移阻害のための方法
US6809077B2 (en) 2001-10-12 2004-10-26 Enanta Pharmaceuticals, Inc. Cyclosporin analogs for the treatment of autoimmune diseases
ATE425178T1 (de) 2001-10-19 2009-03-15 Isotechnika Inc Synthese von cyclosporinanalogen
US7261875B2 (en) * 2001-12-21 2007-08-28 Board Of Regents, The University Of Texas System Dendritic poly (amino acid) carriers and methods of use
FR2836474B1 (fr) * 2002-02-22 2004-12-24 Synt Em Composes, compositions et methode pour le transport des molecules de cyclosporine a travers la barriere hemato-encephalique
US6987090B2 (en) 2002-05-09 2006-01-17 Lg Household & Health Care Ltd. Use of 3-position cyclosporin derivatives for hair growth
DE10223078B4 (de) 2002-05-21 2005-12-22 Novosom Ag Polyampholyte und Verwendung als Polyelektrolytmultischichten
DE10230917A1 (de) 2002-07-09 2004-02-05 Bioleads Gmbh Fredericamycin-Derivate
US7135298B2 (en) 2003-03-26 2006-11-14 The Burnham Institute For Medical Research Screening assay for agents that alter target of Rapamycin activity
ATE419252T1 (de) 2003-10-31 2009-01-15 Cv Therapeutics Inc Antagonisten des a2b-adenosinrezeptors
US7431915B2 (en) * 2003-10-31 2008-10-07 The Regents Of The University Of California Peptides whose uptake by cells is controllable
AU2005238484A1 (en) 2004-04-22 2005-11-10 The Procter & Gamble Company Tri-substituted ureas as cytokine inhibitors
US7259156B2 (en) 2004-05-20 2007-08-21 Kosan Biosciences Incorporated Geldanamycin compounds and method of use
US7226906B2 (en) 2004-08-16 2007-06-05 Array Biopharma, Inc. Cyclosporin analogs for the treatment of immunoregulatory disorders and respiratory diseases
AR050374A1 (es) 2004-08-20 2006-10-18 Wyeth Corp Forma polimorfica de rafampicina
JP2008514701A (ja) 2004-09-29 2008-05-08 エーエムアール テクノロジー インコーポレイテッド シクロスポリンアルキン類似体およびそれらの薬学的使用
US7196161B2 (en) 2004-10-01 2007-03-27 Scynexis Inc. 3-ether and 3-thioether substituted cyclosporin derivatives for the treatment and prevention of hepatitis C infection
US7361636B2 (en) 2004-10-06 2008-04-22 Amr Technology, Inc. Cyclosporin alkynes and their utility as pharmaceutical agents
MX2007007409A (es) 2004-12-20 2007-07-12 Wyeth Corp Derivados de rapamicina y los usos de los mismos en el tratamiento de trastornos neurologicos.
RU2394036C2 (ru) 2004-12-20 2010-07-10 Вайет Аналоги рапамицина и их применение при лечении неврологических, пролиферативных и воспалительных заболеваний
JP2008540571A (ja) 2005-05-12 2008-11-20 ウィスコンシン・アルムニ・リサーチ・ファウンデーション Ｐｉｎ１の遮断は活性化した免疫細胞によるサイトカイン産生を防ぐ

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6030941A (en) * 1996-05-01 2000-02-29 Avi Biopharma, Inc. Polymer composition for delivering substances in living organisms
WO1999030164A1 (fr) * 1997-12-09 1999-06-17 President And Fellows Of Harvard College Procede d'identification de modulateurs de transcription

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