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ES2548030T3 - Moléculas con semividas prolongadas y usos de las mismas - Google Patents

Moléculas con semividas prolongadas y usos de las mismas Download PDF

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Publication number
ES2548030T3
ES2548030T3 ES10783845.0T ES10783845T ES2548030T3 ES 2548030 T3 ES2548030 T3 ES 2548030T3 ES 10783845 T ES10783845 T ES 10783845T ES 2548030 T3 ES2548030 T3 ES 2548030T3
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abd
seq
domain
fragment
amino acid
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Olga Lubman
William Dall'acqua
Herren Wu
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MedImmune LLC
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MedImmune LLC
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
    • C07K14/315—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Streptococcus (G), e.g. Enterococci
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A61K39/39533—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals
    • A61K39/3955—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals against proteinaceous materials, e.g. enzymes, hormones, lymphokines
    • 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
    • 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
    • A61K47/68—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 an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6835—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 an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/52—Constant or Fc region; Isotype
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/94—Stability, e.g. half-life, pH, temperature or enzyme-resistance
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/20—Fusion polypeptide containing a tag with affinity for a non-protein ligand
    • C07K2319/21—Fusion polypeptide containing a tag with affinity for a non-protein ligand containing a His-tag
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/30—Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/31—Fusion polypeptide fusions, other than Fc, for prolonged plasma life, e.g. albumin
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/40—Fusion polypeptide containing a tag for immunodetection, or an epitope for immunisation
    • C07K2319/41—Fusion polypeptide containing a tag for immunodetection, or an epitope for immunisation containing a Myc-tag

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Public Health (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Veterinary Medicine (AREA)
  • Animal Behavior & Ethology (AREA)
  • Epidemiology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Engineering & Computer Science (AREA)
  • Immunology (AREA)
  • Genetics & Genomics (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Molecular Biology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Endocrinology (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Peptides Or Proteins (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

Una molécula que comprende un dominio de unión a albúmina (ABD) de la proteína G de Streptococcus fusionado a un fragmento Fc de IgG de unión a FcRn, en la que la molécula que comprende el ABD fusionado al fragmento Fc de IgG de unión a FcRn tiene una semivida más prolongada que una molécula que comprende el fragmento Fc de IgG de unión a FcRn sin un ABD o que comprende el ABD sin el fragmento Fc de IgG de unión a FcRn

Description

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E10783845
22-09-2015
Dominio de unión a albúmina (ABD)
El ABD se deriva de Streptococcus. Los dominios ABD se describen en el documento Johansson et al. (The Journal of Biological Chemistry, 277: 8114-8120, 2002).
Típicamente, el dominio ABD de la presente invención es pequeño, por ejemplo, aproximadamente 70, 60, 50, 49, 48, 47, 46, 45, 44, 43, 42, 41, 40, 39, 38, 37,36, 35, 34, 33, 32, 31, 30, 28, 27 ó 26 aminoácidos de longitud y puede unirse a albúmina, tal como albúmina de suero humano. Por ejemplo, el dominio ABD puede unirse a albúmina a una afinidad de 1 mM a 1 nM. Por consiguiente, el dominio ABD de la presente invención derivado de una proteína capaz de unirse a albúmina de suero humano (por ejemplo, proteína G estreptocócica) comprenderá al menos esa parte suficiente para unirse a albúmina de suero humano (por ejemplo, SEC ID Nº 1), pero no comprenderá toda la secuencia de aminoácidos de la proteína. El dominio puede incluir, además, un haz de triple hélice tal como un haz de triple hélice levógiro. En ciertas realizaciones, el dominio ABD comprende la SEC ID Nº 1 o la SEC ID Nº 2. En otras realizaciones, el dominio ABD consta de la SEC ID Nº 1 o la SEC ID Nº 2.
La divulgación también incluye variantes de dominios ABD conocidos. Las variantes de ABD pueden contener al menos una alteración de aminoácidos, tal como una sustitución de aminoácidos conservativa en comparación con la secuencia de aminoácidos del ABD de tipo silvestre o puede incluir deleciones de aminoácidos del dominio. Las variantes de ABD pueden tener al menos el 99 %, 98 %, 97 %, 96 %, 95 %, 95 %, 94 %, 93 %, 92 %, 91 %, 90 %, 85 %, 80 %, 75 %, 70 % o 65 % de identidad de secuencia con el ABD de tipo silvestre.
Las variantes de ABD típicamente tienen la misma actividad o sustancialmente la misma actividad que los ABD de tipo silvestre. Por ejemplo, la actividad puede ser la capacidad del dominio para unirse a albúmina, por ejemplo, la variante puede unirse a albúmina con una afinidad (KD) de al menos 1 mM. Las variantes de ABD también incluyen fragmentos de ABD que conservan la capacidad de unirse a albúmina. El fragmento puede ser un dominio ABD que carece, por ejemplo, de 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 o más aminoácidos.
Los ejemplos de ABD de la presente invención incluyen:
G148-GA3: LAEAKVLANRELDKYGVSDYYKNLINNAKTVEGVKALIDEILAALP (SEC ID Nº 1)
ALB8-GA: LKNAKEDAIAELKKAGITSDFYFNAINKAKTVEEVNALKNEILKA (SEC ID Nº 2)
Se apreciará que variantes de G148-GA3 y ALB8-GA pueden tener al menos el 99 %, 98 %, 97 %, 96 %, 95 %, 95 %, 94 %, 93 %, 92 %, 91 %, 90 %, 85 %, 80 %, 75 %, 70 % o 65 % de identidad de secuencia con la SEC ID Nº1 o la SEC ID Nº2 y conservan la capacidad de unirse a albúmina de suero humano.
Las variantes de G148-GA3 y ALB8-GA también pueden incluir ABD que tienen sustituciones, deleciones o inserciones de aminoácidos de uno o más aminoácidos. En un ejemplo, el ABD tiene 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20 o más sustituciones de aminoácidos conservativas.
En un ejemplo, la divulgación incluye una variante de G148-GA3. La variante de G148-GA3 debe tener sustancialmente la misma actividad que G148-GA3 y unirse a albúmina de suero humano con aproximadamente la misma afinidad. Además, la G148-GA3 debe mostrar una conformación de haz de triple hélice levógiro. Residuos críticos para la unión a albúmina están presentes en la hélice 2 e incluyen los residuos S 18, Y20 y Y21 y residuos críticos para la formación del haz de triple hélice levógiro incluyen L12, V33, 1I37 e I40. (Los residuos críticos están en negrita y subrayados a continuación.) Los residuos críticos pueden ser menos tolerantes a sustituciones de aminoácidos no conservativas, por consiguiente, mantenerlos es generalmente preferido. Sin embargo, pueden utilizarse sustituciones conservativas en residuos críticos.
LAEAKVLANRELDKYGVSDYYKNLINNAKTVEGVKALIDEILAALP (SEC ID Nº 1)
De este modo, una variante de G148-GA3 debe conservar los aminoácidos críticos y puede incluir una o más sustituciones de aminoácidos de otra forma a lo largo de la longitud de la SEC ID Nº 1 o ser un fragmento de la SEC ID Nº1.
Variantes del dominio ABD pueden ensayarse para su capacidad de unirse a albúmina de suero humano (por ejemplo tal como se perfila en el ejemplo 2). Las variantes pueden ensayarse, además, para determinar si pueden unirse al FcRn (por ejemplo, tal como se perfila en el ejemplo 3)
La invención incluye, además, las secuencias de ácido nucleico que codifican los ABD de la invención. Debido a la inherente degeneración del código genético, otras secuencias de ADN que codifican sustancialmente la misma o una secuencia de aminoácidos de funcionalidad equivalente, pueden usarse para clonar y expresar el dominio ABD.
Fragmento Fc de IgG de unión a FcRn
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Claims (1)

  1. imagen1
ES10783845.0T 2009-06-01 2010-05-28 Moléculas con semividas prolongadas y usos de las mismas Active ES2548030T3 (es)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US18285809P 2009-06-01 2009-06-01
US182858P 2009-06-01
PCT/US2010/036497 WO2010141329A1 (en) 2009-06-01 2010-05-28 Molecules with extended half-lives and uses thereof

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ES2548030T3 true ES2548030T3 (es) 2015-10-13

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EP (1) EP2437767B1 (es)
DK (1) DK2437767T3 (es)
ES (1) ES2548030T3 (es)
WO (1) WO2010141329A1 (es)

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