Method for stimulating hematopoiesis using tgf-alpha

Abstract

There is disclosed a novel genus of small peptides, much smaller than TGFα, was discovered as having TGFα biological activity and therefore are useful as pharmacologic agents for the same indications as full length TGFα polypeptide. There is further disclosed that TGFα and consequently the genus of small peptides disclosed herein, was found to have therapeutic activity to stimulate hematopoiesis in patients undergoing cytotoxic cancer chemotherapy and to act as a cytoprotective agent to protect a patient undergoing cancer cytotoxic therapy from gastrointestinal (GI) side effects, such as mucositis and otherwise support the barrier function of the GI tract when it is harmed by cytotoxic therapy.

We claim:  
     
         1 . A compound that acts as a TGFα mimetic, comprising at least an 11-membered peptide compound from formula I:  
       N—X 1a -Cys-His-Ser-X 1b —X 2 —X 1a —X 1b —X 1a —X 3 -Cys COOH  I  
       wherein X 1  is independently Val, Gly or Ala, wherein X 2  is Try or Phe, wherein X 3  is Arg or Lys, and wherein the two Cys moieties form a disulfide bond to create an 11-amino acid loop peptide.  
     
     
         2 . The compound of  claim 1  wherein at least one or more of the following seven amino acids are added to the C terminus Cys moiety from formula II:  
       —X 4 -His-X 1c —X 4 —X 5 —X 6 —X 1c   II  
       wherein X 4  is Glu or Asp, wherein X 5  is Leu or Ile, and wherein X 6  is Asp or Glu.  
     
     
         3 . The compound of  claim 1  wherein X 1a  is Val, X 1b  is Gly and X 1c  is Ala.  
     
     
         4 . The compound of  claim 2  wherein X 2  is Tyr, and X 3  is Arg.  
     
     
         5 . The compound of  claim 2  wherein the loop peptide is 13 amino acids in length, wherein X 1a  is Val, X 1b  is Gly, X 1c  is Ala, and X 4  is Gly.  
     
     
         6 . A pharmaceutical composition comprising a loop peptide in a pharmaceutically acceptable carrier, wherein the loop peptide compound comprises at least an 11-membered peptide compound from formula I:  
       N—X 1a -Cys-His-Ser-X 1b —X 2 —X 1a —X 1b —X 1a —X 3 -Cys COOH  I  
       wherein X 1  is independently Val, Gly or Ala, wherein X 2  is Try or Phe, wherein X 3  is Arg or Lys, and wherein the two Cys moieties form a disulfide bond to create an 11-amino acid loop peptide.  
     
     
         7 . The pharmaceutical composition of  claim 6  wherein at least one or more of the following seven amino acids are added to the C terminus Cys moiety from formula II:  
       —X 4 -His-X 1c —X 4 —X 5 —X 6 —X 1c   II  
       wherein X 4  is Glu or Asp, wherein X 5  is Leu or Ile, and wherein X 6  is Asp or Glu.  
     
     
         8 . The pharmaceutical composition of  claim 6  wherein X 1a  is Val, X 1b  is Gly and X 1c  is Ala.  
     
     
         9 . The pharmaceutical composition of  claim 7  wherein X 2  is Tyr, and X 3  is Arg.  
     
     
         10 . The pharmaceutical composition of  claim 7  wherein the loop peptide is 13 amino acids in length wherein X 1a  is Val, X 1b  is Gly, X 1c  is Ala, and X 4  is Gly.  
     
     
         11 . A method for treating a neurodegenerative disease with an pharmaceutically active loop peptide, wherein the loop peptide comprises at least an 11-membered peptide compound from formula I:  
       N—X 1a -Cys-His-Ser-X 1b —X 2 —X 1a —X 1b —X 1a —X 3 -Cys COOH  I  
       wherein X 1  is independently Val, Gly or Ala, wherein X 2  is Try or Phe, wherein X 3  is Arg or Lys, and wherein the two Cys moieties form a disulfide bond to create an 11-amino acid loop peptide.  
     
     
         12 . The method for treating a neurodegenerative disease of  claim 11  wherein at least one or more of the following seven amino acids are added to the C terminus Cys moiety from formula II:  
       —X 4 -His-X 1c —X 4 —X 5 —X 6 —X 1c   II  
       wherein X 4  is Glu or Asp, wherein X 5  is Leu or Ile, and wherein X 6  is Asp or Glu.  
     
     
         13 . The method for treating a neurodegenerative disease of  claim 11  wherein X 1a  is Val, X 1b  is Gly and X 1c  is Ala.  
     
     
         14 . The method for treating a neurodegenerative disease of  claim 12  wherein X 2  is Tyr, and X 3  is Arg.  
     
     
         15 . The method for treating a neurodegenerative disease of  claim 12  wherein the loop peptide is 13 amino acids in length wherein X 1a  is Val, X 1b  is Gly, X 1c  is Ala, and X 4  is Gly.  
     
     
         16 . A method for treating a CNS disease or disorder, wherein the CNS disease or disorder is selected from the group consisting of CNS ischemia, spinal cord injury, MS, and retinal injury, comprising with an pharmaceutically active loop peptide, wherein the loop peptide comprises at least an 11-membered peptide compound from formula I:  
       N—X 1a -Cys-His-Ser-X 1b —X 2 —X 1a —X 1b —X 1a —X 3 -Cys COOH  I  
       wherein X 1  is independently Val, Gly or Ala, wherein X 2  is Try or Phe, wherein X 3  is Arg or Lys, and wherein the two Cys moieties form a disulfide bond to create an 11-amino acid loop peptide.  
     
     
         17 . The method for treating a CNS disease or disorder of  claim 16  wherein at least one or more of the following seven amino acids are added to the C terminus Cys moiety from formula II:  
       —X 4 -His-X 1c —X 4 —X 5 —X 6 —X 1c   II  
       wherein X 4  is Glu or Asp, wherein X 5  is Leu or Ile, and wherein X 6  is Asp or Glu.  
     
     
         18 . The method for treating a CNS disease or disorder of  claim 16  wherein X 1a  is Val, X 1b  is Gly and X 1c  is Ala.  
     
     
         19 . The method for treating a CNS disease or disorder of  claim 17  wherein X 2  is Tyr, and X 3  is Arg.  
     
     
         20 . The method for treating a CNS disease or disorder of  claim 17  wherein the loop peptide is 13 amino acids in length wherein X 1a  is Val, X 1b  is Gly, X 1c  is Ala, and X 4  is Gly.  
     
     
         21 . A method for augmenting hematopoiesis during cytotoxic or immune-suppressing therapy, comprising administering a TGFα polypeptide or a pharmaceutically active loop peptide, or both, wherein the loop peptide comprises at least an 11-membered peptide compound from formula I:  
       N—X 1a -Cys-His-Ser-X 1b —X 2 —X 1a —X 1b —X 1a —X 3 -Cys COOH  I  
       wherein X 1  is independently Val, Gly or Ala, wherein X 2  is Try or Phe, wherein X 3  is Arg or Lys, and wherein the two Cys moieties form a disulfide bond to create an 11-amino acid loop peptide.  
     
     
         22 . The method for augmenting hematopoiesis during cytotoxic or immune-suppressing therapy of  claim 21  wherein at least one or more of the following seven amino acids are added to the C terminus Cys moiety from formula II:  
       —X 4 -His-X 1c —X 4 —X 5 —X 6 —X 1c   II  
       wherein X 4  is Glu or Asp, wherein X 5  is Leu or Ile, and wherein X 6  is Asp or Glu.  
     
     
         23 . The method for augmenting hematopoiesis during cytotoxic or immune-suppressing therapy of  claim 21  wherein X 1a  is Val, X 1b  is Gly and X 1c  is Ala.  
     
     
         24 . The method for augmenting hematopoiesis during cytotoxic or immune-suppressing therapy of  claim 22  wherein X 2  is Tyr, and X 3  is Arg.  
     
     
         25 . The method for augmenting hematopoiesis during cytotoxic or immune-suppressing therapy of  claim 22  wherein the loop peptide is 13 amino acids in length wherein X 1a  is Val, X 1b  is Gly, X 1c  is Ala, and X 4  is Gly.  
     
     
         26 . The method for augmenting hematopoiesis during cytotoxic or immune-suppressing therapy of  claim 21  further comprising administering a second hematopoietic growth factor agent to stimulate more mature hematopoietic precursor cells, wherein the second hematopoietic growth factor is selected from the group consisting of erythropoietin, thrombopoietin, G-CSF (granulocyte colony stimulating factor), and GM-CSF (granulocyte macrophage colony stimulating factor).  
     
     
         27 . A method for treating or preventing mucositis of the gastrointestinal tract during cytotoxic or immune-suppressing therapy, comprising administering a TGFα polypeptide or a pharmaceutically active loop peptide, or both, wherein the loop peptide comprises at least an 11-membered peptide compound from formula I:  
       N—X 1a -Cys-His-Ser-X 1b —X 2 —X 1a —X 1b —X 1a —X 3 -Cys COOH  I  
       wherein X 1  is independently Val, Gly or Ala, wherein X 2  is Try or Phe, wherein X 3  is Arg or Lys, and wherein the two Cys moieties form a disulfide bond to create an 11-amino acid loop peptide.  
     
     
         28 . The method for treating or preventing mucositis of the gastrointestinal tract during cytotoxic or immune-suppressing therapy of  claim 27  wherein at least one or more of the following seven amino acids are added to the C terminus Cys moiety from formula II:  
       —X 4 -His-X 1c —X 4 —X 5 —X 6 —X 1c   II  
       wherein X 4  is Glu or Asp, wherein X 5  is Leu or Ile, and wherein X 6  is Asp or Glu.  
     
     
         29 . The method for treating or preventing mucositis of the gastrointestinal tract during cytotoxic or immune-suppressing therapy of  claim 27  wherein X 1a  is Val, X 1b  is Gly and X 1c  is Ala.  
     
     
         30 . The method for treating or preventing mucositis of the gastrointestinal tract during cytotoxic or immune-suppressing therapy of  claim 28  wherein X 2  is Tyr, and X 3  is Arg.  
     
     
         31 . The method for treating or preventing mucositis of the gastrointestinal tract during cytotoxic or immune-suppressing therapy of  claim 28  wherein the loop peptide is 13 amino acids in length wherein X 1a  is Val, X 1b  is Gly, X 1c  is Ala, and X 4  is Gly.  
     
     
         32 . A bifunctional compound that acts as a TGFα mimetic, comprising a compound from formula III:  
       Loop peptide N-terminus-linker-cyclic C 4 H 8 N 2 -linker-Loop peptide N-terminus  III  
       wherein the linker moiety is designed to link the N-terminus of the Loop peptide to a nitrogen atom of the ring C 4 H 8 N 2  and wherein the “loop peptide” comprises at least an 11-membered peptide compound from formula I:  
       N—X 1a -Cys-His-Ser-X 1b —X 2 —X 1a —X 1b —X 1a —X 3 -Cys COOH  I  
       wherein X 1  is independently Val, Gly or Ala, wherein X 2  is Try or Phe, wherein X 3  is Arg or Lys, and wherein the two Cys moieties form a disulfide bond to create an 11-amino acid loop peptide.  
     
     
         33 . The bifunctional compound that acts as a TGFα mimetic of  claim 32  wherein at least one or more of the following seven amino acids are added to the C terminus Cys moiety from formula II:  
       —X 4 -His-X 1c —X 4 —X 5 —X 6 —X 1c   II  
       wherein X 4  is Glu or Asp, wherein X 5  is Leu or Ile, and wherein X 6  is Asp or Glu.  
     
     
         34 . The bifunctional compound that acts as a TGFα mimetic of  claim 32  wherein X 1a  is Val, X 1b  is Gly and X 1c  is Ala.  
     
     
         35 . The bifunctional compound that acts as a TGFα mimetic of  claim 32  wherein the linker group is independently selected from the group consisting of substituted or unsubstituted C 1-6  alkyl, substituted or unsubstituted C 2-6  alkenyl, substituted or unsubstituted C 1-6  alkoxy, xylenyl, wherein the substitutions are selected from the group consisting of oxo, epoxyl, hydroxyl, chloryl, bromyl, fluoryl, and amino.  
     
     
         36 . The bifunctional compound that acts as a TGFα mimetic of  claim 33  wherein X 2  is Tyr, and X 3  is Arg.  
     
     
         37 . The bifunctional compound that acts as a TGFα mimetic of  claim 33  wherein the loop peptide is 13 amino acids in length wherein X 1a  is Val, X 1b  is Gly, X 1c  is Ala, and X 4  is Gly.  
     
     
         38 . A method for treating inflammatory bowel disease, colitis, and Chron's Disease of the gastrointestinal tract, comprising administering a TGFα polypeptide or a pharmaceutically active loop peptide, or both, wherein the loop peptide comprises at least an 11-membered peptide compound from formula I:  
       N—X 1a -Cys-His-Ser-X 1b —X 2 —X 1a —X 1b —X 1a —X 3 -Cys COOH  I  
       wherein X 1  is independently Val, Gly or Ala, wherein X 2  is Try or Phe, wherein X 3  is Arg or Lys, and wherein the two Cys moieties form a disulfide bond to create an 11-amino acid loop peptide.  
     
     
         39 . The method for treating inflammatory bowel disease, colitis, and Chron's Disease of the gastrointestinal tract of  claim 38  wherein at least one or more of the following seven amino acids are added to the C terminus Cys moiety from formula II:  
       —X 4 -His-X 1c —X 4   13  X 5 —X 6 —X 1c   II  
       wherein X 4  is Glu or Asp, wherein X 5  is Leu or Ile, and wherein X 6  is Asp or Glu.  
     
     
         40 . The method for treating inflammatory bowel disease, colitis, and Chron's Disease of the gastrointestinal tract of  claim 38  wherein X 1a  is Val, X 1b  is Gly and X 1c  is Ala.  
     
     
         41 . The method for treating inflammatory bowel disease, colitis, and Chron's Disease of the gastrointestinal tract of  claim 39  wherein X 2  is Tyr, and X 3  is Arg.  
     
     
         42 . The method for treating inflammatory bowel disease, colitis, and Chron's Disease of the gastrointestinal tract of  claim 39  wherein the loop peptide is 13 amino acids in length wherein X 1a  is Val, X 1b  is Gly, X 1c  is Ala, and X 4  is Gly.  
     
     
         43 . A method for treating an inflammatory reaction of autoimmune diseases, comprising administering a TGFα polypeptide or a pharmaceutically active loop peptide, or both, wherein the loop peptide comprises at least an 11-membered peptide compound from formula I:  
       N—X 1a -Cys-His-Ser-X 1b —X 2 —X 1a —X 1b —X 1a —X 3 -Cys COOH  I  
       wherein X 1  is independently Val, Gly or Ala, wherein X 2  is Try or Phe, wherein X 3  is Arg or Lys, and wherein the two Cys moieties form a disulfide bond to create an 11-amino acid loop peptide.  
     
     
         44 . The method for treating an inflammatory reaction of autoimmune diseases of  claim 43  wherein the autoimmune diseases are selected from the group consisting of Type II (Juvenile) Diabetes, rheumatoid arthritis, lupus, and multiple sclerosis.  
     
     
         45 . The method for treating an inflammatory reaction of autoimmune diseases of  claim 43  wherein at least one or more of the following seven amino acids are added to the C terminus Cys moiety from formula II:  
       —X 4 -His-X 1c —X 4 —X 5 —X 6 —X 1c   II  
       wherein X 4  is Glu or Asp, wherein X 5  is Leu or Ile, and wherein X 6  is Asp or Glu.  
     
     
         46 . The method for treating an inflammatory reaction of autoimmune diseases of  claim 43  wherein X 1a  is Val, X 1b  is Gly and X 1c  is Ala.  
     
     
         47 . The method for treating an inflammatory reaction of autoimmune diseases of  claim 45  wherein X 2  is Tyr, and X 3  is Arg.  
     
     
         48 . The method for treating an inflammatory reaction of autoimmune diseases of  claim 45  wherein the loop peptide is 13 amino acids in length wherein X 1a  is Val, X 1b  is Gly, X 1c  is Ala, and X 4  is Gly.