Cytokine derived treatment with reduced vascular leak syndrome

Abstract

The present invention relates to a pharmaceutical composition for treating a cancer or an infection in a subject by administrating an amount of an IL-15 derivative conjugate so as to induce a proliferation of natural killer cells (NK cells) which is the same or higher than the one obtained with high dose of interleukin-2 (HDIL-2); eventually associated with a pharmaceutically acceptable carrier.

What is claimed is: 
     
         1 . A method for treating a cancer, an infection, or an immunodeficiency disorder in a human, comprising administering a pharmaceutical composition comprising a conjugate to a subject, wherein the conjugate is administered in an effective amount as to induce a proliferation of natural killer cells (NK cells) which is the same or higher than the one obtained with High Dose of interleukin-2 (HDIL-2), wherein said conjugate comprises:
 a) a polypeptide comprising the amino acid sequence of interleukin 15 or derivatives thereof having an amino acid sequence having a percentage of identity of at least 92.5% with the amino acid sequence of SEQ ID NO:3, and   b) a polypeptide comprising the amino acid sequence of the sushi domain of IL-15Rα or derivatives thereof having an amino acid sequence having a percentage of identity of at least 92% with an amino acid sequence selected in the group consisting of SEQ ID NO:8, SEQ ID NO:9 and SEQ ID NO:12;   optionally associated with a pharmaceutically acceptable carrier, wherein the effective amount of said conjugate is between 24 and 2400 pmol/kg, preferably between 28 and 800 pmol/kg.   
     
     
         2 . The method of  claim 1 , wherein the effective amount of said conjugate is between 0.6 and 60 μg/kg, preferably between 0.7 and 20 μg/kg. 
     
     
         3 . The method of  claim 1 , wherein said interleukin 15 derivative has at least 10% of the activity of human interleukin-15 on the proliferation induction of kit225 cell line, preferably at least 25% and more preferably at least 50%, and/or
 has an amino acid sequence having a percentage of identity of at least 98.5% with the amino acid sequence of SEQ ID NO:3, preferably of at least 99%.   
     
     
         4 . The method of  claim 1 , wherein said derivative of the sushi domain of the IL-15Rα has at least 10%, preferably at least 25% and more preferably at least 50% of the binding activity of the sushi domain of human IL-15Rα, and/or
 has an amino acid sequence having a percentage of identity of at least 96% with an amino acid sequence in the group consisting of SEQ ID NO:8, SEQ ID NO:9 and SEQ ID NO:12, preferably of at least 98%. 
 
     
     
         5 . The method of  claim 1 , wherein said conjugate is administrated in an amount inducing a proliferation of CD8 T cells higher than the one obtained with HDIL-2. 
     
     
         6 . The method of  claim 1 , wherein said composition is for treating an advanced (TNM grade IV) or a metastatic cancer and said administrated amount of conjugate induce a proliferation of NK cells which is higher than the one obtained with HDIL-2. 
     
     
         7 . The method of  claim 6 , wherein said conjugate is administrated
 (I) in an amount inducing a proliferation of NK cells, which is at least 20% higher than the one obtained with HDIL-2; preferably at least 25% higher, and still preferably at least 30% higher than the one obtained with HDIL-2; and/or   (II) in an amount inducing a proliferation of CD8+ T cells, which is at least 20% higher than the one obtained with HDIL-2; preferably at least 25% higher; and still preferably at least 30% higher than the one obtained with HDIL-2.   
     
     
         8 . The method of  claim 1 , wherein said composition is for treating anon-metastatic cancer, preferably a TNM grade I, II or III cancer, or an infection, and said administrated amount of conjugate induces a proliferation of NK cells, which is the same or higher than the one obtained with HDIL-2. 
     
     
         9 . The method of  claim 8 , wherein
 (I) said conjugate is administrated in an amount inducing a proliferation of natural killer cells (NK cells) which is the same or at the maximum 50 or 25% higher than the one obtained with HDIL-2; preferably the same or at the maximum 20% higher; and still preferably the same or at the maximum 10% higher than the one obtained with HDIL-2; and/or   (II) said conjugate is administrated in an amount inducing a proliferation of CD8 +  T cells, which is the same or at the maximum 200% higher than the one obtained with HDIL-2; preferably the same or at the maximum 150% higher; and still preferably the same or at the maximum 100% higher than the one obtained with HDIL-2; and/or   (III) wherein the conjugate is administrated in an amount corresponding to a blood concentration comprised between 40 fmol/ml and 12 pmol/ml (1 ng/ml and 300 ng/ml), preferably between 80 fmol/ml and 12 pmol/ml (2 ng/ml and 300 ng/ml), and still preferably between 0.16 and 4 pmol/ml (4 and 100 ng/ml).   
     
     
         10 . The method of  claim 1 , wherein the administrated amount of conjugate induces a proliferation of Treg cells (FoxP3 + CD4 + CD25 high ) which is less than the one obtained with HDIL-2, wherein the administrated amount of conjugate preferably induces a proliferation of Treg cells which is at least 5% less than the one obtained with HDIL-2; preferably at least 10 or 20% less; and still preferably at least 50% less than the one obtained with HDIL-2. 
     
     
         11 . The method of  claim 1 , wherein
 (I) the administrated amount corresponds to a ratio of induced percentage of proliferating NK cells on induced percentage of proliferating Treg cells which is at least 25% higher than the one obtained with HDIL-2; preferably at least 50% higher; and still preferably at least 75% higher than the one obtained with HDIL-2; and/or   (II) the administrated amount of conjugate corresponds to a ratio of induced percentage of proliferating CD8 T cells on the induced percentage of proliferating Treg cells which is at least 25% higher than the one obtained with HDIL-2; preferably at least 50% higher; and still preferably at least 75% higher than the one obtained with HDIL-2.   
     
     
         12 . The method of  claim 1 , wherein
 (I) the polypeptides a) and b) of the conjugate are covalently linked in a fusion protein; and/or   (II) said conjugate comprises the amino acid sequence of the interleukin 15 or derivatives thereof in a C-terminal position relative to the amino acid sequence of the sushi domain of the IL-15Rα or derivatives thereof; and/or   (III) the amino acid sequence of the interleukin 15 or derivatives thereof and the amino acid sequence of the sushi domain of the IL-15Rα or derivatives are separated by a linker amino acid sequence having a length of 5-30 amino acids, said linker comprising near neutral amino acids selected in the group comprising Gly (G), Asn (N), Ser (S), Thr (T), Ala (A), Leu (L), and Gln (Q).   
     
     
         13 . The method of  claim 1 , wherein
 (I) the administrated amount corresponds to a daily administration amount; and/or   (II) said composition is administrated parenterally, preferably intravenously.   
     
     
         14 . An in vitro method for determining the therapeutically efficient amount of a conjugate to be administrated to a subject suffering from a cancer, from an infection or from an immunodeficiency disorder, said method comprising the steps of:
 i) contacting peripheral blood mononucleated cells (PBMCs) from said subject with increasing amounts of a conjugate in culture conditions enabling the proliferation of said PBMCs;   ii) contacting other PBMCs from said subject with High Dose of interleukin-2 (HDIL-2) in culture conditions enabling the proliferation of said PBMCs; and   iv) selecting a therapeutically efficient amount of conjugate, said therapeutically efficient amount inducing a proliferation of NK cells of said PBMCs which is the same or higher than the one obtained with HDIL-2;   wherein said conjugate comprises:   a) a polypeptide comprising the amino acid sequence of interleukin 15 or derivatives thereof having an amino acid sequence having a percentage of identity of at least 92.5% with the amino acid sequence of SEQ ID NO:3, and   b) a polypeptide comprising the amino acid sequence of the sushi domain of IL-15Rα or derivatives thereof having an amino acid sequence having a percentage of identity of at least 92% with an amino acid sequence selected in the group consisting of SEQ ID NO:8, SEQ ID NO:9 and SEQ ID NO:12.   
     
     
         15 . The method of  claim 14 , wherein
 (I) said therapeutically efficient amount of conjugate induces a proliferation of CD8 T cells of said PBMCs which is the same or higher than the one obtained with HDIL-2; and/or   (II) said therapeutically efficient amount of conjugate corresponds to a ratio of the induced percentage of proliferating NK cells and/or of CD8 T cells on the induced percentage of proliferating Treg cells which is at least 25% higher than the one obtained with HDIL-2; preferably at least 50% higher; and still preferably at least 75% higher than the one obtained with HDIL-2; and/or   (III) the increasing amounts of conjugate correspond to concentration of conjugate comprised between 4 fmol/ml and 120 pmol/ml (0,1 and 3,000 ng/ml), preferably between 40 fmol/ml and 80 pmol/ml (1 and 2,000 ng/ml), and still preferably between 80 fmol/ml and 40 pmol/ml (2 and 1,000 ng/ml); and/or   (IV) said method of the invention further comprises the step of:
 iii) contacting peripheral blood mononucleated cells (PBMCs) from said subject with increasing equimolar amounts of IL-15 as compared to the conjugate in culture conditions enabling the proliferation of said PBMCs.