Composition for improving skin conditions comprising a fragment of human heat shock protein 90a as an active ingredient

Abstract

Liposomal and/or nano-liposomal encapsulated peptides of HSP90a, HPf polypeptide (115 aa) and novel polypeptides HPfΔC1 (101 aa) and HPfΔC2 (87 aa), and methods for manufacturing/preparing and using the compositions, are disclosed. Chimeric fusion proteins that include HSP90a, HPf, HPfAC, HPfAC2 polypeptide, or combinations thereof, are presented. Transformed cell lines and expression vectors capable of expressing the chimeric fusion proteins, are provided. Methods for producing large amounts of recombinant HSP90a, HPf polypeptide, HPfΔC1 or HPfΔC2 polypeptide, using expression vectors and transformed cell lines, are described. Topical and other delivery form preparations, including microneedle preparations, and methods for using the preparations for improving skin conditions (atopic dermatitis, wrinkles, skin elasticity, dark spots (over pigmentation), overall skin rejuvenation, skin ageing) and other therapeutic (anti-cancer, anti-ALS, anti-Huntington's disease, obesity) and cosmeceutical uses are presented. Wound healing preparations with the Hsp90a and related peptides are disclosed.

What is claimed is: 
     
         1 . A chimeric construct encoding a fusion protein comprising a nucleic acid sequence encoding HSP90a, HPf polypeptide, HFfΔC1 or HPfΔC2 polypeptide, or a fragment thereof, and a nucleic acid sequence encoding a fusion partner peptide. 
     
     
         2 . The chimeric construct of  claim 1  wherein the fusion partner peptide is theoredoxin A, maltose binding protein (MBP) or human growth hormone (hGH). 
     
     
         3 . The chimeric construct of  claim 2  further comprising a protein cleavage enzyme recognition site located between the nucleic acid sequence encoding the HPf peptide and the nucleic acid sequence encoding the fusion partner peptide. 
     
     
         4 . The chimeric construct of  claim 1  wherein the HPf polypeptide has a nucleic acid sequence of SEQ ID. NO. 1. 
     
     
         5 . A transformed cell line transformed to express a HPf-fusion partner chimeric protein, said cell line comprising a TOP 10 cell line, an RZ4500 cell line, a BL21(DE3)pLyS cell line or a RosettaBlue (DE3) cell line. 
     
     
         6 . The transformed cell line of  claim 5  wherein the HPf-fusion partner chimeric protein is TRX(TEVc)-HPf fusion protein, MBP(TEVc)-HPf fusion protein, or TRX(NGc)-HPf fusion protein. 
     
     
         7 . A method for preparing a nano-liposomal encapsulated HSP90a, HPf, HFfΔC1 or HPfΔC2 polypeptide composition, said method comprising:
 (a) dissolving a phospholipid capable of forming a liposome with an HSP90a, HPf, HFfΔC1 or HPfΔC2 polypeptide, in a buffered aqueous solution of salt, said phospholipid comprising yellow yolk lecithin or soybean lecithin; and 
 (b) passing the aqueous solution through a high-pressure homogenizer while gradually increasing the content of the phospholipid and the pressure of the high-pressure homogenizer as the number of the passages increases to provide an HSP90a, HPf, HFfΔC1 or HPfΔC2 polypeptide-containing nano-liposomal encapsulated HPf polypeptide composition. 
 
     
     
         8 . The method of  claim 7  wherein the polypeptide is HSP90a. 
     
     
         9 . A method for treating a skin condition, wherein the skin condition is atopic dermatitis, wrinkles, dark spots, skin elasticity or skin aging, comprising:
 providing a composition comprising a concentration of about 100 ng/ml to about 1 mg/ml of an Hsp90a, HPf, HFfΔC1 or HPfΔC2 polypeptide, or a combination thereof, to a skin area demonstrating atopic dermatitis, wrinkles, dark spots, skin elasticity or skin aging; and   reducing the skin condition at the skin area.   
     
     
         10 . A method for reducing subcutaneous fat accumulation comprising:
 providing a composition comprising a concentration of about 100 ng/ml to about 1 mg/ml of an Hsp90a, HPf, HFfΔC1, HPfΔC2 polypeptide, or a combination thereof, to an area presenting subcutaneous fat accumulation; and   reducing subcutaneous fat in the area on which the composition is applied.   
     
     
         11 . The method of  claim 10  wherein the composition comprises a patch or body wrap including a microneedle formulation of the Hsp90a, HPf, HFfΔC1, HPfΔC2, or a combination thereof. 
     
     
         12 . A method for inhibiting synuclein-induced toxicity related neurological disease, comprising:
 providing a composition comprising a concentration of about 100 ng/ml to about 1 mg/ml of an HPf polypeptide, HPf polypeptide, HFfΔC1, HPfΔC2, or fragment thereof, to a patient having a synuclein-induced toxicity related neurological disease; and   inhibiting neurological degeneration from synuclein-induced toxicity.   
     
     
         13 . The method of  claim 12  wherein the neurological disease is Huntington's disease or Amyotrophic lateral sclerosis. 
     
     
         14 . A dissolvable microneedle formulation comprising HSP90a, HPf polypeptide, HPfΔC1, HPfΔC2, or a combination thereof. 
     
     
         15 . The dissolvable microneedle formulation of  claim 14  wherein the HPf polypeptide has a sequence of SEQ ID NO 1, the HPfΔC1 has a sequence of SEQ ID NO 20 and the HPfΔC2 has a sequence of SEQ ID NO 21. 
     
     
         16 . The dissolvable microneedle formulation of  claim 14  comprising a patch, body wrap or facial mask. 
     
     
         17 . A method for treating a neurodegenerative disease associated with neurotoxic tau accumulation comprising:
 providing a patient having a neurodegenerative disease with a therapeutic agent comprising a peptide of HSP90a, HPf polypeptide, HPfΔC1, HPfΔC2, or a combination thereof; and   reducing neurotoxic tau species accumulation,   
       wherein generation of neurotoxic tau species is inhibited. 
     
     
         18 . The method of  claim 17  wherein the therapeutic agent comprises a fusion peptide, wherein said fusion peptide comprises a peptide of HSP90a, HPf, HPfΔC1, HPfΔC2, or a combination thereof and a growth factor peptide or protein. 
     
     
         19 . The method of  claim 18  wherein the growth factor is human growth factor. 
     
     
         20 . The method of  claim 17  wherein the neurodegenerative disease is Alzheimers disease.