Patent Application
20240408148
The present invention relates generally to the field of umbilical cord derived compositions, and more particularly to non-immunogenic compositions for dermal application to a subject in need thereof in which the compositions are derived from fresh human umbilical cords.
Compositions derived from human umbilical cords have various uses in the medical field. For example, these advantageous uses may include harvesting stem cells therefrom for the potential treatment of various blood diseases, cancers, and immune system disorders. When preparing compositions derived from human umbilical cords, the umbilical cord tissue is often subjected to harsh mechanical and enzymatic processing conditions in which specific cells (e.g., stem cells) may be isolated from the umbilical cord tissue, expanded/cultured, and cryopreserved, thus drastically altering the initial, endogenous cellular and extracellular profile of the umbilical cord tissue. Furthermore and either before, during, and/or after processing these umbilical cord isolates, exogenous additives, including various growth factors/cytokines such as interferon alpha (INF-α), are included within these isolates, which further alter these isolates when compared to the initial umbilical cord tissue. These alterations may further decrease efficacy in a desired treatment due to loss of the original, endogenous cellular and/or extracellular profile of the initial umbilical cord tissue.
In view of the above, it is an object of the invention to provide compositions derived from human umbilical cord(s) that mimic, include, and/or retain an extracellular profile similar to the endogenous profile of a human umbilical cord (e.g., in vivo), especially when compared with various previously mentioned umbilical cord isolates. The compositions described herein are prepared with fresh human umbilical cord (harvested and processed within 48 to 72 hours of extraction from the human subject) and, unlike the prior art compositions, are advantageously not subjected to biochemical and/or enzymatic digestion, which results in the compositions including and/or retaining a significant proportion of the extracellular profile (of a human umbilical cord in vivo).
In certain aspects, disclosed is an aqueous non-immunogenic composition for topical, subcutaneous, or intradermal use in a human subject in need thereof. In certain aspects, the aqueous non-immunogenic composition comprises an aqueous human umbilical cord filtrate obtained from human umbilical cord. In certain aspects, the aqueous non-immunogenic composition consists of an aqueous human umbilical cord filtrate obtained from human umbilical cord. In certain aspects, the aqueous non-immunogenic composition consists essentially of an aqueous human umbilical cord filtrate obtained from human umbilical cord.
In certain aspects, the aqueous human umbilical cord filtrate is a solution in which no settling, separation, and/or precipitation is observed after twelve months, twenty-four months, up to 60 months or more while being stored at −20° C. or −80° C. In certain aspects, the aqueous human umbilical cord filtrate is a solution in which no settling, separation, and/or precipitation is observed after ten days, twenty days, thirty days, up to sixty days or more while being stored between 4° C. and 8° C. In certain aspects and when preparing the aqueous human umbilical cord filtrate, no exogenous enzymes are introduced therein, which avoids exogenous enzymatic degradation/digestion. In certain aspects, the aqueous human umbilical cord filtrate is obtained by filtering ground human umbilical cord. In certain aspects, the aqueous non-immunogenic composition has particulates less than 100 μm. In certain aspects, the human umbilical cord is double-filtered to obtain aqueous human umbilical cord filtrate. In certain aspects the human umbilical cord is triple-filtered to obtain human umbilical cord filtrate. In certain aspects, the aqueous non-immunogenic composition has particulates less than 50 μm. In certain aspects, the aqueous non-immunogenic composition has particulates less than 35 μm. In certain aspects, the aqueous non-immunogenic composition has particulates less than 10 μm. In certain aspects, the aqueous non-immunogenic composition is sterile. In certain aspects, the aqueous non-immunogenic composition is acellular.
In certain aspects, the aqueous human umbilical cord filtrate comprises at least one of: acellular Wharton's jelly, exosomes, endogenous growth factors, vascular endothelial growth factor receptor-1 (VEGFR1) at a concentration of 1.0×102 pg/mL to 2.5×103 pg/mL, hepatocyte growth factor (HGF) at a concentration of 1.38×102 pg/mL to 1.42×104 μg/mL, interleukin antagonists (interleukin-1 receptor antagonist (IL-1ra)) at a concentration of 3.18×102 pg/mL to 5.15×104 pg/mL, platelet derived growth factor-BB (PDGF-BB) at a concentration of 1.5×101 pg/mL to 1.58×103 pg/mL, basic fibroblast growth factor (bFGF) at a concentration of 3.5×101 pg/ml to 2.07×103 pg/ml, endogenous hyaluronic acid (HA) at a concentration of 1.29×104 μg/mL to 3.5×108 pg/mL, or any combination thereof.
In certain aspects, the aqueous human umbilical cord filtrate comprises at least two of: acellular Wharton's jelly, exosomes, endogenous growth factors, vascular endothelial growth factor receptor-1 (VEGFR1) at a concentration of 1.0×102 pg/mL to 2.5×103 pg/mL, hepatocyte growth factor (HGF) at a concentration of 1.38×102 pg/mL to 1.42×104 pg/mL, interleukin antagonists (interleukin-1 receptor antagonist (IL-1ra)) at a concentration of 3.18×102 pg/mL to 5.15×104 pg/mL, platelet derived growth factor-BB (PDGF-BB) at a concentration of 1.5×101 pg/mL to 1.58×103 pg/mL, basic fibroblast growth factor (bFGF) at a concentration of 3.5×101 pg/ml to 2.07×103 pg/ml, endogenous hyaluronic acid (HA) at a concentration of 1.29×104 pg/mL to 3.5×108 pg/mL, or any combination thereof.
In certain aspects, the aqueous human umbilical cord filtrate comprises at least three of: acellular Wharton's jelly, exosomes, endogenous growth factors, vascular endothelial growth factor receptor-1 (VEGFR1) at a concentration of 1.0×102 pg/mL to 2.5×103 pg/mL, hepatocyte growth factor (HGF) at a concentration of 1.38×102 pg/mL to 1.42×104 μg/mL, interleukin antagonists (interleukin-1 receptor antagonist (IL-1ra)) at a concentration of 3.18×102 pg/mL to 5.15×104 pg/mL, platelet derived growth factor-BB (PDGF-BB) at a concentration of 1.5×101 pg/mL to 1.58×103 pg/mL, basic fibroblast growth factor (bFGF) at a concentration of 3.5×101 pg/ml to 2.07×103 μg/ml, endogenous hyaluronic acid (HA) at a concentration of 1.29×104 pg/mL to 3.5×108 pg/mL, or any combination thereof.
In certain aspects, the aqueous human umbilical cord filtrate comprises at least four of: acellular Wharton's jelly, exosomes, endogenous growth factors, vascular endothelial growth factor receptor-1 (VEGFR1) at a concentration of 1.0×102 pg/mL to 2.5×103 pg/mL, hepatocyte growth factor (HGF) at a concentration of 1.38×102 pg/mL to 1.42×104 pg/mL, interleukin antagonists (interleukin-1 receptor antagonist (IL-1ra)) at a concentration of 3.18×102 pg/mL to 5.15×104 pg/mL, platelet derived growth factor-BB (PDGF-BB) at a concentration of 1.5×101 pg/mL to 1.58×103 pg/mL, basic fibroblast growth factor (bFGF) at a concentration of 3.5×101 pg/ml to 2.07×103 μg/ml, endogenous hyaluronic acid (HA) at a concentration of 1.29×104 pg/mL to 3.5×108 pg/mL, or any combination thereof.
In certain aspects, the aqueous human umbilical cord filtrate comprises at least five of: acellular Wharton's jelly, exosomes, endogenous growth factors, vascular endothelial growth factor receptor-1 (VEGFR1) at a concentration of 1.0×102 pg/mL to 2.5×103 pg/mL, hepatocyte growth factor (HGF) at a concentration of 1.38×102 pg/mL to 1.42×104 μg/mL, interleukin antagonists (interleukin-1 receptor antagonist (IL-1ra)) at a concentration of 3.18×102 pg/mL to 5.15×104 pg/mL, platelet derived growth factor-BB (PDGF-BB) at a concentration of 1.5×101 pg/mL to 1.58×103 pg/mL, basic fibroblast growth factor (bFGF) at a concentration of 3.5×101 pg/ml to 2.07×103 pg/ml, endogenous hyaluronic acid (HA) at a concentration of 1.29×104 pg/mL to 3.5×108 pg/mL, or any combination thereof.
In certain aspects, the aqueous human umbilical cord filtrate comprises at least six of: acellular Wharton's jelly, exosomes, endogenous growth factors, vascular endothelial growth factor receptor-1 (VEGFR1) at a concentration of 1.0×102 pg/mL to 2.5×103 pg/mL, hepatocyte growth factor (HGF) at a concentration of 1.38×102 pg/mL to 1.42×104 μg/mL, interleukin antagonists (interleukin-1 receptor antagonist (IL-1ra)) at a concentration of 3.18×102 pg/mL to 5.15×104 pg/mL, platelet derived growth factor-BB (PDGF-BB) at a concentration of 1.5×101 pg/mL to 1.58×103 pg/mL, basic fibroblast growth factor (bFGF) at a concentration of 3.5×101 pg/ml to 2.07×103 pg/ml, endogenous hyaluronic acid (HA) at a concentration of 1.29×104 pg/mL to 3.5×108 pg/mL, or any combination thereof.
In certain aspects, the aqueous human umbilical cord filtrate comprises at least seven of: acellular Wharton's jelly, exosomes, endogenous growth factors, vascular endothelial growth factor receptor-1 (VEGFR1) at a concentration of 1.0×102 pg/mL to 2.5×103 pg/mL, hepatocyte growth factor (HGF) at a concentration of 1.38×102 pg/mL to 1.42×104 μg/mL, interleukin antagonists (interleukin-1 receptor antagonist (IL-1ra)) at a concentration of 3.18×102 pg/mL to 5.15×104 pg/mL, platelet derived growth factor-BB (PDGF-BB) at a concentration of 1.5×101 pg/mL to 1.58×103 pg/mL, basic fibroblast growth factor (bFGF) at a concentration of 3.5×101 pg/ml to 2.07×103 pg/ml, endogenous hyaluronic acid (HA) at a concentration of 1.29×104 pg/mL to 3.5×108 pg/mL, or any combination thereof.
In certain aspects, the aqueous human umbilical cord filtrate comprises at least eight of: acellular Wharton's jelly, exosomes, endogenous growth factors, vascular endothelial growth factor receptor-1 (VEGFR1) at a concentration of 1.0×102 pg/mL to 2.5×103 pg/mL, hepatocyte growth factor (HGF) at a concentration of 1.38×102 pg/mL to 1.42×104 μg/mL, interleukin antagonists (interleukin-1 receptor antagonist (IL-1ra)) at a concentration of 3.18×102 pg/mL to 5.15×104 pg/mL, platelet derived growth factor-BB (PDGF-BB) at a concentration of 1.5×101 pg/mL to 1.58×103 pg/mL, basic fibroblast growth factor (bFGF) at a concentration of 3.5×101 pg/ml to 2.07×103 pg/ml, endogenous hyaluronic acid (HA) at a concentration of 1.29×104 pg/mL to 3.5×108 pg/mL, or any combination thereof.
In certain aspects, the composition further includes one or more excipients that facilitate epidermal penetration, dermal penetration, extracellular matrix penetration, or a combination thereof. In certain aspects, the one or more excipients comprises water, glycerol/glycerides, alcohols/alkanols (e.g. ethanol, glycols, methanol, isopropyls, butanols, pentanols), surfectants (nonionic, cationic, and anionic) (e.g. Laureth-9), detergents (e.g. Tween 20, Brij 35), Fatty acids (e.g. oleic acid, laurate, myristic acid, capric acid), terpenes and derivatives (e.g. menthol, thymol, carvacrol, menthone, cineole), amides (including urea and its derivatives), fatty alcohols (e.g. lauryl, stearyl, and oleyl) and their derivatives, essential oils (e.g. aloe vera, cumin, tea tree oils, Copaiba oil, eucalyptus oil, cinanamon oil, Mentha haplocalyx essential oil), sulfoxides (e.g. dimethyl sulfoxide (DMSO), chealators (e.g. ethylenediaminetetraacetic acid (EDTA), salicylic acid), Glycols (e.g. Propylene glycol (monocaprylate, isostearate, monolaurate)), polyoxylglycerides, cyclodextrin, sodium glycocholate and related cholates, saponins, trihydroxy salts (bile salts), pyrrolidones, esters, esters sulfoxides and their derivatives, hydrocarbons, 1-dodecylazacycloheptan-2-one and its analogs, oleodendrimers, ionic liquids, and deep eutectic solvents, fusidic acid derivatives, caprylate, phospholipids, nanoparticles, ethosomes, dendrimers, liposomes, laurocapram, azone, oxazolidinones, and n-disubstituted amino acetates.
In certain aspects, the composition is formulated as a cream. In certain aspects, the composition is formulated as a solution. In certain aspects, the composition is formulated as an ointment. In certain aspects, the composition is formulated as a gel. The composition may be formulated with glyceryl dibehenate, glyceryl distearate, polyglyceryl-3 dioleate, ethylcellulose, behenoyl polyoxyl-8 glycerides, glyceryl monostearate, refined soybean oil (glyceryl palmitostearate) or hard fat to create a cream, ointment or gel.
In certain aspects, the aqueous human umbilical cord filtrate further includes an isotonic solution. In certain aspects, the isotonic solution is phosphate buffered saline (1×PBS), lactated ringers (sodium chloride 6 g/L, sodium lactate 3.1 g/L, potassium chloride 0.3 g/L, and calcium chloride 0.2 g/L at pH 6.5), isotonic saline (0.9 wt % sodium chloride), Plasmalyte® (sodium chloride 5.26 g/L, potassium chloride 0.37 g/L, magnesium chloride hexahydrate 0.30 g/L, sodium acetate trihydrate 3.68 g/L, sodium gluconate 5.02 g/L at pH 7.4), or Normosol® (sodium chloride 5.26 g/L, KCl 0.37 g/L, magnesium chloride 0.30 g/L, sodium acetate anhydrous 2.22 g/L, sodium gluconate 5.02 g/L at pH 7.4). In certain aspects, the aqueous human umbilical cord filtrate further includes amniotic fluid.
In certain aspects, the filtrates mentioned immediately above may be further combined with an isotonic solution (a diluent) that may further dilute growth factor concentrations to a desired range. In certain aspects and when an isotonic solution is included therein, the composition comprises at least one of acellular Wharton's jelly, exosomes, endogenous growth factors, vascular endothelial growth factor receptor (VEGFR1) at a concentration ranging from 1.23×102 pg/mL to 1.9×103 pg/mL, hepatocyte growth factor (HGF) at a concentration ranging from 1.38×102 pg/mL to 1.0×103 pg/mL, interleukin antagonists (interleukin-1 receptor antagonist (IL-1ra)) at a concentration ranging from 3.18×102 pg/mL to 3.95×103 pg/mL, platelet derived growth factor-BB (PDGF-BB) at a concentration ranging from 1.9×101 pg/mL to 2.65×102 pg/mL, basic fibroblast growth factor (bFGF) at a concentration of 3.5×101 pg/mL to 1.5×102 pg/mL, endogenous hyaluronan (HA) at a concentration ranging from 1.29×104 pg/mL to 1.5×108 pg/mL, or any combination thereof.
In certain aspects and when an isotonic solution is included therein, the composition comprises at least two of acellular Wharton's jelly, exosomes, endogenous growth factors, vascular endothelial growth factor receptor (VEGFR1) at a concentration ranging from 1.23×102 pg/mL to 1.9×103 pg/mL, hepatocyte growth factor (HGF) at a concentration ranging from 1.38×102 pg/mL to 1.0×103 pg/mL, interleukin antagonists (interleukin-1 receptor antagonist (IL-1ra)) at a concentration ranging from 3.18×102 pg/mL to 3.95×103 pg/mL, platelet derived growth factor-BB (PDGF-BB) at a concentration ranging from 1.9×101 pg/mL to 2.65×102 pg/mL, basic fibroblast growth factor (bFGF) at a concentration of 3.5×101 pg/mL to 1.5×102 pg/mL, endogenous hyaluronan (HA) at a concentration ranging from 1.29×104 pg/mL to 1.5×108 pg/mL, or any combination thereof.
In certain aspects and when an isotonic solution is included therein, the composition comprises at least three of acellular Wharton's jelly, exosomes, endogenous growth factors, vascular endothelial growth factor receptor (VEGFR1) at a concentration ranging from 1.23×102 pg/mL to 1.9×103 pg/mL, hepatocyte growth factor (HGF) at a concentration ranging from 1.38×102 pg/mL to 1.0×103 pg/mL, interleukin antagonists (interleukin-1 receptor antagonist (IL-1ra)) at a concentration ranging from 3.18×102 pg/mL to 3.95×103 pg/mL, platelet derived growth factor-BB (PDGF-BB) at a concentration ranging from 1.9×101 pg/mL to 2.65×102 pg/mL, basic fibroblast growth factor (bFGF) at a concentration of 3.5×101 pg/mL to 1.5×102 pg/mL, endogenous hyaluronan (HA) at a concentration ranging from 1.29×104 pg/mL to 1.5×108 pg/mL, or any combination thereof.
In certain aspects and when an isotonic solution is included therein, the composition comprises at least four of acellular Wharton's jelly, exosomes, endogenous growth factors, vascular endothelial growth factor receptor (VEGFR1) at a concentration ranging from 1.23×102 pg/mL to 1.9×103 pg/mL, hepatocyte growth factor (HGF) at a concentration ranging from 1.38×102 pg/mL to 1.0×103 pg/mL, interleukin antagonists (interleukin-1 receptor antagonist (IL-1ra)) at a concentration ranging from 3.18×102 pg/mL to 3.95×103 pg/mL, platelet derived growth factor-BB (PDGF-BB) at a concentration ranging from 1.9×101 pg/mL to 2.65×102 pg/mL, basic fibroblast growth factor (bFGF) at a concentration of 3.5×101 pg/mL to 1.5×102 pg/mL, endogenous hyaluronan (HA) at a concentration ranging from 1.29×104 pg/mL to 1.5×108 pg/mL, or any combination thereof.
In certain aspects and when an isotonic solution is included therein, the composition comprises at least five of acellular Wharton's jelly, exosomes, endogenous growth factors, vascular endothelial growth factor receptor (VEGFR1) at a concentration ranging from 1.23×102 pg/mL to 1.9×103 pg/mL, hepatocyte growth factor (HGF) at a concentration ranging from 1.38×102 pg/mL to 1.0×103 pg/mL, interleukin antagonists (interleukin-1 receptor antagonist (IL-1ra)) at a concentration ranging from 3.18×102 pg/mL to 3.95×103 pg/mL, platelet derived growth factor-BB (PDGF-BB) at a concentration ranging from 1.9×101 pg/mL to 2.65×102 pg/mL, basic fibroblast growth factor (bFGF) at a concentration of 3.5×101 pg/mL to 1.5×102 pg/mL, endogenous hyaluronan (HA) at a concentration ranging from 1.29×104 pg/mL to 1.5×108 pg/mL, or any combination thereof.
In certain aspects and when an isotonic solution is included therein, the composition comprises at least six of acellular Wharton's jelly, exosomes, endogenous growth factors, vascular endothelial growth factor receptor (VEGFR1) at a concentration ranging from 1.23×102 pg/mL to 1.9×103 pg/mL, hepatocyte growth factor (HGF) at a concentration ranging from 1.38×102 pg/mL to 1.0×103 pg/mL, interleukin antagonists (interleukin-1 receptor antagonist (IL-1ra)) at a concentration ranging from 3.18×102 pg/mL to 3.95×103 pg/mL, platelet derived growth factor-BB (PDGF-BB) at a concentration ranging from 1.9×101 pg/mL to 2.65×102 pg/mL, basic fibroblast growth factor (bFGF) at a concentration of 3.5×101 pg/mL to 1.5×102 pg/mL, endogenous hyaluronan (HA) at a concentration ranging from 1.29×104 pg/mL to 1.5×108 pg/mL, or any combination thereof.
In certain aspects and when an isotonic solution is included therein, the composition comprises at least seven of acellular Wharton's jelly, exosomes, endogenous growth factors, vascular endothelial growth factor receptor (VEGFR1) at a concentration ranging from 1.23×102 pg/mL to 1.9×103 pg/mL, hepatocyte growth factor (HGF) at a concentration ranging from 1.38×102 pg/mL to 1.0×103 pg/mL, interleukin antagonists (interleukin-1 receptor antagonist (IL-1ra)) at a concentration ranging from 3.18×102 pg/mL to 3.95×103 pg/mL, platelet derived growth factor-BB (PDGF-BB) at a concentration ranging from 1.9×101 pg/mL to 2.65×102 pg/mL, basic fibroblast growth factor (bFGF) at a concentration of 3.5×101 pg/mL to 1.5×102 pg/mL, endogenous hyaluronan (HA) at a concentration ranging from 1.29×104 pg/mL to 1.5×108 pg/mL, or any combination thereof.
In certain aspects and when an isotonic solution is included therein, the composition comprises at least eight of acellular Wharton's jelly, exosomes, endogenous growth factors, vascular endothelial growth factor receptor (VEGFR1) at a concentration ranging from 1.23×102 pg/mL to 1.9×103 pg/mL, hepatocyte growth factor (HGF) at a concentration ranging from 1.38×102 pg/mL to 1.0×103 pg/mL, interleukin antagonists (interleukin-1 receptor antagonist (IL-1ra)) at a concentration ranging from 3.18×102 pg/mL to 3.95×103 pg/mL, platelet derived growth factor-BB (PDGF-BB) at a concentration ranging from 1.9×101 pg/mL to 2.65×102 pg/mL, basic fibroblast growth factor (bFGF) at a concentration of 3.5×101 pg/mL to 1.5×102 pg/mL, endogenous hyaluronan (HA) at a concentration ranging from 1.29×104 pg/mL to 1.5×108 pg/mL, or any combination thereof.
In certain aspects, the composition further includes an effective amount of exogenous hyaluronic acid to improve and/or restore appearance and function to at least one of the skin, mucosa, epidermis, dermis, extracellular matrix, or any combination thereof. In certain aspects, the exogenous hyaluronic acid is present in the composition at a concentration of 0.8 mg/ml to 30 mg/ml. In other aspects, the composition has a concentration of about 1% to about 50% exogenous hyaluronic acid. In other aspects, the composition has a concentration of about 5 mg/ml to 20 mg/ml. In other aspects, the composition has a concentration of about 10 mg/ml to 15 mg/ml.
In certain aspects, the composition is configured intradermal use in a human subject in need thereof as a sterile composition. In this aspect the composition is preferably a viscous aqueous composition having a sufficient viscosity for an intradermal injection into a dermis with minimal pain and/or discomfort to the subject and sufficient thickness and consistency to improve and/or restore appearance and function to at least one of the skin, mucosa, epidermis, dermis, extracellular matrix, or any combination thereof. In certain aspects, the composition is configured for subcutaneous use in a human subject in need thereof as a sterile composition. In this aspect the composition is preferably a viscous aqueous composition having a sufficient viscosity for a subcutaneous injection underneath the dermis of the skin with minimal pain and/or discomfort to the subject and sufficient thickness and consistency to improve and/or restore appearance and function to at least one of the skin, mucosa, epidermis, dermis, extracellular matrix, or any combination thereof
In certain aspects, the composition is configured for topical use in a human subject in need thereof as a topically applied composition. In this aspect the composition is preferably a viscous aqueous composition having a sufficient viscosity for topical application onto the epidermis with minimal pain and/or discomfort to the subject and sufficient thickness and consistency to improve and/or restore appearance and function to at least one of the skin, mucosa, epidermis, dermis, extracellular matrix, or any combination thereof. In certain aspects, also disclosed is a method of administering the aqueous, non-immunogenic composition for topical, subcutaneous, or intradermal use to a human subject in need thereof. The method includes step (1): applying an effective amount of the composition on skin, on mucosa, on and/or in the extracellular matrix of the human subject in need thereof to improve and/or restore appearance and function to at least one of the skin, mucosa, epidermis, dermis, extracellular matrix, or any combination thereof by treating and/or reducing symptoms of inflammation in the subject. In certain aspects, the composition is sterile. In certain aspects, the composition further comprises an effective amount of exogenous hyaluronic acid to improve and/or restore appearance and function to at least one of the skin, mucosa, epidermis, dermis, extracellular matrix, or any combination thereof. In certain aspects, the exogenous hyaluronic acid is present in the composition at a concentration of 0.8 mg/ml to 30 mg/ml. In other aspects, the composition has a concentration of about 1% to about 50% exogenous hyaluronic acid. In other aspects, the composition has a concentration of about 5 mg/ml to 20 mg/ml. In other aspects, the composition has a concentration of about 10 mg/ml to 15 mg/ml.
In certain aspects, step (1) includes applying the composition via intradermal injection. This method comprises sterilely injecting the composition into dermis just below the epidermis in the human subject in need thereof. In certain aspects, step (1) includes applying the composition via subcutaneous injection. This method comprises sterilely injecting the composition into the subcutis layer underneath the dermis. In certain aspects, step (1) includes applying the composition topically. This method includes applying the composition onto the epidermis.
In certain aspects, the method further includes, prior to step (1), administering a physical dermal treatment. In certain aspects, the physical dermal treatment comprises microneedling. In certain aspects, the physical dermal treatment comprises dermabrasion. In certain aspects, the physical dermal treatment comprises microdermabrasion.
In certain aspects, applying an effective amount of the composition to at least one of the skin, mucosa, epidermis, dermis, extracellular matrix, or any combination thereof of the human subject in need thereof to improve and/or restore appearance and function to at least one of the skin, mucosa, epidermis, dermis, extracellular matrix, or any combination thereof is repeated at predetermined time intervals. In certain aspects, step (1) is repeated daily. In certain aspects, step (1) is repeated weekly. In certain aspects, step (1) is repeated monthly. In certain aspects, step (1) is repeated bi-weekly. In certain aspects, step (1) is repeated semi-weekly. In certain aspects, step (1) is repeated bi-monthly. In certain aspects, step (1) is repeated semi-monthly.
In certain aspects, the composition is administered to the human subject in need thereof to provide treatment or prevention for signs of aging, acne, scarring, psoriasis, eczema, and/or skin inflammation. Dermal conditions, treatments, or insults that result in breaking the epidermal barrier, dermal layer, and/or hypodermis by puncture, scratch, abrasion, laceration, tear, burn (chemical or physical) or bruising. (e.g. microneedling, dermabrasion, chemical peel, laser resurfacing).
Embodiments of the invention can include one or more or any combination of the above features and configurations.
Additional features, aspects and advantages of the invention will be set forth in the detailed description which follows, and in part will be readily apparent to those skilled in the art from that description or recognized by practicing the invention as described herein. It is to be understood that both the foregoing general description and the following detailed description present various embodiments of the invention, and are intended to provide an overview or framework for understanding the nature and character of the invention as it is claimed. The accompanying drawings are included to provide a further understanding of the invention, and are incorporated in and constitute a part of this specification.
These and other features, aspects and advantages of the present invention are better understood when the following detailed description of the invention is read with reference to the accompanying drawings, in which:
The present invention will now be described more fully hereinafter with reference to the accompanying drawings in which exemplary embodiments of the invention are shown. However, the invention may be embodied in many different forms and should not be construed as limited to the representative embodiments set forth herein. The exemplary embodiments are provided so that this disclosure will be both thorough and complete, and will fully convey the scope of the invention and enable one of ordinary skill in the art to make, use and practice the invention. Like reference numbers refer to like elements throughout the various drawings. Moreover, in this specification and in the claims that follow, reference will be made to a number of terms that shall be defined to have the following meanings:
It must be noted that, as used in the specification and the appended claims, the singular forms “a,” “an” and “the” include plural referents unless the context clearly dictates otherwise.
Concentrations, amounts, and other numerical data may be expressed or presented herein in a range format. It is to be understood that such a range format is used merely for convenience and brevity and thus should be interpreted flexibly to include not only the numerical values explicitly recited as the limits of the range, but also to include all the individual numerical values or sub-ranges encompassed within the ranges as if each numerical value and sub-range is explicitly recited. As an illustration, a numerical range of “about 1 to 5” should be interpreted to include not only the explicitly recited values of about 1 to about 5, but also include individual values and sub-ranges within the indicated range. Thus, included in this numerical range are individual values such as 2, 3, and 4 and sub-ranges such as from 1-3, from 2-4, and from 3-5, etc. as well as 1, 2, 3, 4, and 5, individually. The same principle applies to ranges reciting only one numerical value as a minimum or a maximum. Furthermore, such an interpretation should apply regardless of the breadth of the range or the characteristics being described.
The compositions and methods described herein can comprise, consist of, or consist essentially of the essential elements and limitations described herein, as well as any additional or optional ingredients, components, or limitations described herein.
Disclosed herein are compositions derived from human umbilical cord(s) that retain an extracellular profile similar to the endogenous profile of a human umbilical cord, for example, in vivo, especially when compared with various previously mentioned umbilical cord isolates. These compositions are prepared with fresh human umbilical cord (harvested and processed within 48 to 72 hours of extraction from the human subject) and, unlike compositions in the prior art, are advantageously not subjected to biochemical and/or enzymatic digestion, which results in the compositions including and/or retaining a significant portion of the extracellular profile (when compared to the endogenous profile of a human umbilical cord in vivo). Moreover because of the ease and convenience of administering these compositions (e.g., point of use preparation and use within a dental office, medical office, or emergency room, or use by the patient in their own home) and because of the non-immunogenic characteristics of these compositions, these compositions may be used for numerous different medical purposes and medical procedures, which include, but are not limited to, topical, subcutaneous, or intradermal use in a human subject in need thereof to improve and/or restore endogenous extracellular matrix function in at least one of the skin, mucosa, epidermis, dermis, hypodermis, extracellular matrix, or any combination thereof; improve and/or restore endogenous collagen function in at least one of the skin, mucosa, epidermis, dermis, hypodermis, extracellular matrix, or any combination thereof, to treat and/or reduce symptoms of inflammation in the subject, or any combination thereof.
Disclosed herein are aqueous non-immunogenic compositions (e.g., compositions) that include an aqueous human umbilical cord filtrate which may be configured for topical, subcutaneous, or intradermal use. In certain aspects and when preparing the composition no exogenous enzymes are introduced therein, which avoids exogenous enzymatic degradation/digestion and further ensures that these compositions have an improved endogenous extracellular profile (similar to human umbilical cord in vivo) especially when compared to conventional compositions utilizing umbilical cord tissues and/or cells derived therefrom.
Hyaluronic acid is the main component of the extracellular matrix and other human connective tissue and plays a number of structural roles in vivo. Endogenous hyaluronan and sulfated glycosaminoglycans (sGAGs), found within the composition described herein, increases the tensile strength of the extracellular matrix within the skin. Additionally, hyaluronic acid may trigger intracellular events that lead to an increase in cell migration and proliferation. Application of hyaluronic acid alone has been shown to improve smoothness and hydration in the skin and improve skin flexibility. An additional property of hyaluronic acid is its ability to absorb water, or hygroscopicity. This property is desirable for topical, subcutaneous, or intradermal use as it would draw water to the epidermis, dermis, and/or extracellular matrix and improve the overall appearance and overall feel of the skin, as well as promote healing.
The aqueous human umbilical cord filtrate, of the composition, is prepared, preferably from human umbilical cord, via one or more separation steps (e.g., filtration steps). The human umbilical cord filtrate preferably includes acellular Wharton's jelly, exosomes, endogenous growth factors, vascular endothelial growth factor receptor 1 (VEGFR1), hepatocyte growth factor (HGF), interleukin antagonists (IL-1ra), platelet derived growth factor-BB (PDGF-BB), basic fibroblast growth factor (bFGF), endogenous hyaluronan (HA) or a combination thereof therein, which advantageously restores the structure and function of the dermal extracellular matrix within a subject when the disclosed compositions are used for their desired purpose. As shown in
In certain aspects, the aqueous human umbilical cord filtrate may include particles that remain from a human umbilical cord tissue therein that are less than 100 μm in diameter, preferably less than 50 μm in diameter, more preferably less than 35 μm in diameter, even more preferably less than 10 μm in diameter. Moreover, aqueous human umbilical cord filtrate is a solution in which no settling, separation, and/or precipitation is observed after one month, two months, three months, four months, five months, six months, or more while being stored. In certain aspects and due to the preparation steps of these compositions as disclosed immediately below as well as in
In certain aspects, exogenous hyaluronic acid may be added to the aqueous human umbilical cord filtrate. In certain aspects, the exogenous hyaluronic acid is present in the composition at a concentration of 0.8 mg/ml to 30 mg/ml. In other aspects, the composition has a concentration of about 1% to about 50% exogenous hyaluronic acid. In other aspects, the composition has a concentration of about 5 mg/ml to 20 mg/ml. In other aspects, the composition has a concentration of about 10 mg/ml to 15 mg/ml.
Hyaluronic acid is also referred to as hyaluronan or hyaluronate; these terms are used interchangeably throughout this specification. Hyaluronic acid is a glycosaminoglycan consisting of repeating units of D-glucoronic acid and N-acetyl-D-glucosamine. The hyaluronic acid used herein may be in salt form or non-salt form. Salt forms of hyaluronic acid include sodium hyaluronate, potassium hyaluronate, calcium hyaluronate, and magnesium hyaluronate. In some aspects, the hyaluronic acid used herein may be obtained from biofermentation in bacteria, including but not limited to: Enterococcus faecalis, Streptococcus zooepidemicus, Escherichia coli, Agrobacterium sp., Lactococcus lactis, and Bacillus subtilis. In other aspects, recombinant hyaluronic acid production is the source of the exogenous hyaluronic acid used herein. One example of recombinant hyaluronic acid production includes the expression hyaluronic acid synthase and UDP-glucose dehydrogenase in a host bacteria to produce large quantities of hyaluronic acid in a fed-batch culture process. In other aspects, the hyaluronic acid used herein may be obtained via extraction from animal tissues, including but not limited to: rooster combs, bovine synovial fluid, and vitreous humor of cattle. In some aspects, hyaluronic acid may be purchased from commercial sources, including but not limited to: Kewpie, Awa Biopharm, Dongchen Group, Fufeng Group, Focus Chem, and Bloomage Biotech.
The exogenous hyaluronic acid used herein may be in a variety of molecular weights. The term “molecular weight” may refer to both the weight-average molecular weight and the number-average molecular weight. In some aspects, the hyaluronic acid used herein may have a molecular weight of about 20 kDa to about 20,000 kDa. In some aspects, the hyaluronic acid used herein may have a molecular weight of about 20 kDa to 7000 kDa.
In some aspects, the exogenous hyaluronic acid within the aqueous human umbilical cord filtrate is cross-linked. Hyaluronic acid may be cross-linked using a variety of cross-linking agents including, but not limited to, 1,4-butanediol diglycidyl ether (BDDE), poly (ethylene glycol) diglycidyl ether (PEGDE), pentaerythritol tetraglycidyl ether (PETGE), divinyl sulfone, 1,2-bis(2,3-epoxypropoxy)ethylene (EGDGE), 1,2,7,8-diepoxyoctane (DEO), (phenylenebis-(ethyl)-carbodiimde, 1,6-hexamethylenebis (ethylcarbodiimide), adipic dihydrazide (ADH), bis(sulfosuccinimdyl) suberate (BS), hexamethylenediiamine (HMDA), and 1-(2,3-epoxypropyl)-2,3-epoxycyclohexane. The degree of crosslinking, as used herein, is defined as the percent of free hyaluronic acid (non-cross-linked hyaluronic acid). In some aspects, the exogenous hyaluronic acid is heavily cross-linked, with a low percentage of free hyaluronic acid, such as 5%-25%. In some aspects, the exogenous hyaluronic acid may be mildly cross-linked, about 26%-74% free hyaluronic acid. In some aspects, the exogenous hyaluronic acid may be lightly cross-linked, with a high percentage of free hyaluronic acid-75%-95% free hyaluronic acid. In some aspects, the hyaluronic acid is non cross-linked or 100% free hyaluronic acid. The degree of cross-linking within the hyaluronic acid increases the half-life of hyaluronic acid within the body, and thus longer therapeutic effects may be observed.
As further alluded to above, the compositions may be used within humans for numerous different purposes and procedures, which include, but are not limited to, topical, subcutaneous, or intradermal use. In this aspect, it is important to maintain sterility of the aqueous human umbilical cord filtrate. It should be noted that the aqueous human umbilical cord filtrate is non-immunogenic, and thus, should induce very little immune response within a subject when used for its desired purpose. However, sterility should be maintained such that contaminants (e.g., viral contaminants, bacterial contaminants, chemical contaminants, etc.) are not introduced into the composition that may induce an immune response and/or cause infection when the composition is placed in or on a subject.
The composition may be administered to treat and/or prevent acne in a human subject in need thereof. Additionally, or alternatively, the composition may be administered to treat and/or prevent scarring in a human subject in need thereof. In certain aspects, the composition may be administered to prevent and/or reverse signs of aging in a human subject in need thereof. In certain aspects, the composition is administered in combination with a second therapy. In certain aspects, the composition may be administered to treat psoriasis in a human subject in need thereof. In certain aspects, the composition may be administered to treat eczema in a human subject in need thereof. In certain aspects, the composition may be administered to reduce skin inflammation in a human subject in need thereof.
The aqueous, non-immunogenic composition may be formulated with a pharmaceutical or food grade excipient, or carrier, the biological system or entity can tolerate including, but not limited to: dimethyl sulfoxide (DMSO). A pharmaceutical or food grade carrier or excipient, refers to a carrier or an excipient that is useful in preparing a composition that is generally safe and non-toxic, while providing a stable formulation at the desired concentration, consistency, and form. Pharmaceutical or food grade carriers or excipients may also be added to facilitate penetration into the epidermis, dermis, and/or extracellular matrix. Pharmaceutical grade and food grade excipients are known to those skilled in the art. These most typically would be standard carriers for administration to humans and animals, including solutions such as buffered solutions at physiological pH. Nonaqueous carriers, such as fixed oils, vegetable oils such as olive oil, flax seed oil and sesame oil, propylene glycol, and polyethylene glycol can also be used to further enhance delivery and efficacy of the components of the aqueous, non-immunogenic composition. Excipients can also contain minor amounts of additives, such as substances that enhance isotonicity and chemical stability. The aqueous, non-immunogenic composition may be formulated as a gel, an ointment, a cream, or a solution. Other additives include viscosity enhancing agents, such as sodium carboxymethylcellulose, sorbitol, dextran, xanthan, lecithin, beta glucan, hyaluronic acid, and glycerin.
In certain aspects, the one or more excipients comprises water, glycerol/glycerides, alcohols/alkanols (e.g. ethanol, glycols, methanol, isopropyls, butanols, pentanols), surfectants (nonionic, cationic, and anionic) (e.g. Laureth-9), detergents (e.g. Tween 20, Brij 35), Fatty acids (e.g. oleic acid, laurate, myristic acid, capric acid), terpenes and derivatives (e.g. menthol, thymol, carvacrol, menthone, cineole), amides (including urea and its derivatives), fatty alcohols (e.g. lauryl, stearyl, and oleyl) and their derivatives, essential oils (e.g. aloe vera, cumin, tea tree oils, Copaiba oil, eucalyptus oil, cinanamon oil, Mentha haplocalyx essential oil), sulfoxides (e.g. dimethyl sulfoxide (DMSO), chealators (e.g. ethylenediaminetetraacetic acid (EDTA), salicylic acid), Glycols (e.g. Propylene glycol (monocaprylate, isostearate, monolaurate)), polyoxylglycerides, cyclodextrin, sodium glycocholate and related cholates, saponins, trihydroxy salts (bile salts), pyrrolidones, esters, esters sulfoxides and their derivatives, hydrocarbons, 1-dodecylazacycloheptan-2-one and its analogs, oleodendrimers, ionic liquids, and deep eutectic solvents, fusidic acid derivatives, caprylate, phospholipids, nanoparticles, ethosomes, dendrimers, liposomes, laurocapram, azone, oxazolidinones, and n-disubstituted amino acetates.
In certain aspects, the compositions are configured topical application in a human subject. In this aspect, the resulting composition is preferably a fluid having a sufficient consistency to improve and/or restore appearance and function to at least one of the skin, mucosa, epidermis, dermis, hypodermis, extracellular matrix, or combinations thereof. The composition configured for topical application may be in the form of an ointment, a cream, a gel, or a solution. In certain aspects, the compositions are configured for subcutaneous injection in a human subject. In this aspect, the resulting composition is preferably a fluid having a sufficient consistency improve and/or restore appearance and function to at least one of the skin, mucosa, epidermis, dermis, hypodermis, extracellular matrix, or combinations thereof. The compositions may be injected into the subcutis layer beneath the dermis, of a subjects face, chest, neck, back or essentially anywhere the subject's skin requires treatment. In this aspect, the resulting composition is preferably in the form of a solution.
In certain aspects, the compositions are configured for intradermal injection in a human subject. In this aspect, the resulting composition is preferably a fluid having a sufficient consistency improve and/or restore appearance and function to at least one of the skin, mucosa, epidermis, dermis, hypodermis, extracellular matrix, or combinations thereof. The compositions may be injected into the dermis of a subjects face, chest, neck, back, or essentially anywhere the subject's skin requires treatment. In this aspect, the resulting composition is in the form of a solution.
As shown in
After completing step (a), step (b) occurs. Step (b) includes placing <80 grams of umbilical cord into a container having a predetermined volume (e.g., 300 mL to 1000 mL, preferably 500 mL) of isotonic solution in which the isotonic solution is preferably phosphate buffered saline (PBS) (i.e., 1×PBS) (or alternatively one of lactated ringers (sodium chloride 6 g/L, sodium lactate 3.1 g/L, potassium chloride 0.3 g/L, and CaCl 0.2 g/L at pH 6.5), isotonic saline (0.9 wt % sodium chloride), Plasmalyte® (sodium chloride 5.26 g/L, KCl 0.37 g/L, magnesium chloride hexahydrate 0.30 g/L, sodium acetate trihydrate 3.68 g/L, sodium gluconate 5.02 g/L at pH 7.4), Normosol® (sodium chloride 5.26 g/L, KCl 0.37 g/L, magnesium chloride 0.30 g/L, sodium acetate anhydrous 2.22 g/L, sodium gluconate 5.02 g/L at pH 7.4)). Placing the container onto a stir plate and placing a stir bar within the container (containing the PBS and umbilical cord) therein and stirring (medium to high speed) the umbilical cord within the isotonic solution for 5 to 15 minutes to wash the umbilical cord portions. Next, washing step (b) is repeated one to five times by decanting the “used” isotonic solution and pouring new isotonic solution into the container at a predetermined volume (e.g., 300 mL to 1000 mL, preferably 500 mL) to again wash the umbilical cord. Either before step (a), during step (a), after step (b), or during step (b) further determining whether any blood clots and/or blood pool(s)/pooling are present in the human umbilical cord and/or umbilical cord portions, and if so, removing these blood clots via suction or other mechanical removal means (e.g., scalpel, gauze and forceps) to further ensure that the presence of any immunogenic components (e.g., hemoglobin and/or heme associated components from the umbilical cord donor) are minimized in the end resulting composition. During these washing steps, it is imperative to maintain an aseptic and/or sterile work environment to prevent and/or reduce introduction of any contaminants while making the composition.
Upon concluding step (b), step (c) is performed in which the washed umbilical cord is transferred to a grinding and/or mincing apparatus such as those disclosed in U.S. D716,601 “Tissue Mincing Tool” and/or U.S. Pat. No. 8,967,512 “Systems And Methods For Processing Cells”, which are incorporated by reference herein in their entirety, and a predetermined volume (e.g., 75 mL to 125 mL, preferably 100 mL) of the isotonic solution) is added to the apparatus. The washed umbilical cord is subsequently subjected to grinding and/or mincing by the grinding/mincing tool with the head of the grinding/mincing tool rotating at a range of 40 to 200 revolutions per minute (RPM) until the umbilical cord has been fully ground (or as close to fully ground as possible) thereby forming ground human umbilical cord tissue. During this grinding/mincing step, it is imperative to maintain a sterile work environment to prevent and/or reduce introduction of any contaminants while making the composition. In certain aspects, the grinding/mincing tool may be directly connected to an apparatus (i.e., a closed system environment as disclosed, for example, in U.S. Pat. No. 8,967,512) to further conduct steps (d) and/or (e) discussed below and to further maintain sterility and/or minimize the introduction of any contaminants while making the composition. Alternatively, steps (d) and/or (e) may be conducted in an open system/laboratory environment.
Upon concluding step (c), step (d) is performed in which the ground/minced human umbilical cord tissue of step (c) is separated into a solid retentate and an aqueous human umbilical cord supernatant. This initial separation step may occur via a filtration process (either positive or negative pressure). For example, the minced/ground human umbilical cord tissue (of step (c) and included within a predetermined volume (e.g., 75 mL to 125 mL, preferably 100 mL) of the isotonic solution) may be placed directly on a filter having a desired porosity (e.g., 200 μm or 150 μm or 100 μm such as either a qualitative grade or quantitative grade mesh or net filter) and then force (either positive or negative pressure) may or may not be applied such that a solid retentate (solids having a size above 200 μm or 150 μm or 100 μm) remain on the filter while an aqueous human umbilical cord supernatant (having any solids therein that are less than (200 μm or 150 μm or 100 μm) are passed through the filter. The filtration step generally takes 15 seconds to 2 minutes. Again, it is imperative to maintain a sterile and/or aseptic work environment to prevent and/or reduce introduction of any contaminants throughout step (d).
Upon concluding step (d), optional step (e) may be performed on the aqueous human umbilical cord supernatant. Step (e) preferably includes a plurality of filtration steps including: (i) filtering the aqueous human umbilical cord supernatant through a first filter having a porosity ranging from 30 μm to 40 μm thereby forming a second human umbilical cord supernatant; (ii) filtering the second human umbilical cord supernatant through a second filter having a porosity ranging from 10 μm to 25 μm thereby forming a third human umbilical cord supernatant; and (iii) filtering the third human umbilical cord supernatant through a third filter having a porosity ranging from 4 μm to 10 μm thereby forming the aqueous human umbilical cord filtrate. In certain aspects, the force applied is a negative pressure (vacuum) and preferred because such negative pressure is less likely to damage the filter and lead to subsequent quality control issues with the resulting compositions disclosed herein. The aqueous human umbilical cord filtrate preferably includes acellular Wharton's jelly, exosomes, endogenous growth factors, VEGFR1, HGF, interleukin antagonists (IL-1ra), bFGF, PDGF-BB, endogenous hyaluronan, or any combination thereof. Each filtration step generally takes 15 seconds to 2 minutes at 1-5 psi vacuum to complete. Moreover, the resulting aqueous human umbilical cord filtrate from the above mentioned filtration steps is a solution in which no settling, separation, and/or precipitation is observed after one month, two months, three months, four months, five months, six months, twelve months, twenty-four months, sixty months, or more while being stored. Upon filtering the human umbilical cord supernatant through a filter having a porosity of 10 μm or less, the solution is free from cells, resulting in an acellular supernatant. Therefore, in some aspects, the composition described herein is acellular. In certain aspects, the aqueous human umbilical cord filtrate is sterile, and the aqueous human umbilical cord filtrate is non-immunogenic.
Upon concluding step (d) and/or (e), optional step (f) may be performed where the human umbilical cord filtrate is diluted with an isotonic solution or amniotic fluid, thereby forming a composition standardized to a known factor (e.g. original umbilical cord weight, average growth factor content, etc.) Alternatively, the composition may be diluted with an isotonic solution or amniotic fluid just prior to administration to a human subject in need thereof.
Upon concluding step (d) and/or (e), and either concurrently with (and/or either before or during step (e) and/or (f)), optional step (g) is performed that the solid retentate of step (d) is further processed into a micronized human umbilical cord composition by subjecting the solid retentate to a dehydration (lyophilization), freeze drying, milling and/or (cryomilling), process configured to yield particles (polydisperse particles) having sizes ranging from greater than 1 μm to 300 μm, preferably greater than 1 μm to 100 μm, and more preferably greater than 1 μm to 50 μm and more preferably greater than to than 1 μm to 35 μm. In certain aspects, step (e) is a cryomilling process (as described, for example, US20160287749, US20170203004, and U.S. patent Ser. No. 10/105,398, which are each incorporated by reference in their entirety herein) in which the solid retentate of step (d) is dehydrated and placed into a liquid nitrogen cooled cryomill chamber and subjected to grinding therein, thereby forming the micronized human umbilical cord composition having particle sizes ranging from greater than 1 μm to less than 300 μm, preferably greater than 1 μm to 100 μm, more preferably greater than 1 μm to 50 μm, and even more preferably from greater than 1 μm to 35 μm. The micronized human umbilical cord composition comprises collagen, fibronectin, endogenous hyaluronan, elastins, or any combination thereof. The micronized human umbilical cord may be saved for another application such as combining with the aqueous human umbilical cord filtrate of step (d), (e) or (f) to prepare a two-part composition for other envisioned therapeutic applications.
In certain aspects, after step (d), (e) and/or (f) is performed, exogenous hyaluronic acid is added to the aqueous human umbilical cord filtrate prior to step (h) below. In some aspects, heavily cross-linked hyaluronic acid is added to the aqueous human umbilical cord filtrate. In some aspects, mildly cross-linked hyaluronic acid is added to the aqueous human umbilical cord filtrate. In other aspects, lightly cross-linked hyaluronic acid is added to the aqueous human umbilical cord filtrate. In some aspects, non-cross-linked hyaluronic acid is added to the aqueous human umbilical cord filtrate. In certain aspects, exogenous hyaluronic acid is present in the composition at a concentration of 0.8 mg/ml to 30 mg/ml. In other aspects, the composition has a concentration of about 1% to about 50% exogenous hyaluronic acid. In other aspects, the composition has a concentration of about 5 mg/ml to 20 mg/ml. In other aspects, the composition has a concentration of about 10 mg/ml to 15 mg/ml.
In some aspects, the exogenous hyaluronic acid is added to the aqueous human umbilical cord filtrate as a solid. In other aspects, the exogenous hyaluronic acid is added to the aqueous human umbilical cord filtrate as an aqueous solution.
In certain after step (d), (e) and/or (f) is performed, excipients and/or other additives may be added to the to the aqueous human umbilical cord filtrate prior to step (h) below. A pharmaceutical or food grade carrier or excipient may be added to the composition to provide the desired concentration of component in the composition, desired consistency of the composition, and/or to facilitate penetration in the epidermis, dermis, and/or extracellular matrix.
After concluding step (d), (e) and/or (f), step (h) may be performed by placing and sealing the aqueous human umbilical cord filtrate (of step (d), (e) or (f)) in a sterile container for subsequent use, wherein the aqueous human umbilical cord filtrate is sterile.
Without wishing to be bound by theory, it is envisioned that the compositions disclosed herein may be particularly useful for topical, subcutaneous, or intradermal use, and would advantageously produce very little immunogenic response due to the composition's non-immunogenic characteristics/properties.
The skin is the body's physical barrier against mechanical damage, UV-damage, and chemical damage. Human skin is made up of the epidermis, the dermis, and the hypodermis. The epidermis is composed primarily of keratinocytes, melanocytes, and Langerhans' cells; each having a specific role within the skin. The dermis, which provides a solid support for the epidermis, consists primarily of fibroblasts, leukocytes, and mast cells. The extracellular matrix of the epidermis and dermis contains collagen, elastin, glycosaminoglycans, proteoglycans, fibronectin, glycoproteins, and many other components that, among other functions, provide structural integrity to the skin.
Cosmetic treatments, dysfunctions, and changes in the components of the extracellular matrix of the skin can interfere with tissue integrity and cell performance. This may lead to skin aging, skin atrophy, damaged and/or wounded skin, eczema, psoriasis, inflammation, infection, and other skin conditions. The components of the composition described herein for topical, subcutaneous, or intradermal use can restore structure and function to the extracellular matrix. The extracellular matrix provides support and lubrication for the cells of the dermis and epidermis for proper function. Among the many beneficial components of the aqueous human umbilical cord filtrate, and the composition described herein, are glycosaminoglycans including hyaluronic acid, proteoglycans, IL-1ra, which provides cell growth signaling, anti-inflammatory effects, and provide structure to the extracellular matrix in the at least one of the skin, mucosa, epidermis, dermis, hypodermis or any combination thereof providing a therapeutic or cosmetic effect.
Glycosaminoglycans (GAGs) play an important role in the extracellular matrix. GAGs play a crucial role in skin physiology by regulating keratinocyte proliferation and differentiation, inflammatory processes, and extracellular matrix composition and quality. Particularly important GAGs in the skin include chondroitin sulfate, dermatan sulfate, keratin sulfate, and heparin sulfate.
Hyaluronic acid, a glycosaminoglycan found endogenously in the composition described herein, and in some aspects added exogenously to the composition, is involved in many biological functions, including but not limited to: cell differentiation, embryological development, inflammation, wound healing, hydration and viscoelasticity. Hyaluronic acid has been shown to be an effective at reducing inflammation, regenerating skin, and decreasing the appearance of aging. Hyaluronic acid is an important structural component of the ECM, forming a scaffold for macroproteins. Hyaluronic acid is hygroscopic and promotes tissue hydration and contributes to osmotic balance which stabilizes the extracellular matrix. Hyaluronic acid is widely used in the cosmetics/cosmeceutical industry to increase skin moisture, skin elasticity, and improve the overall health of skin. Additionally, hyaluronic acid forms an occlusive layer on the skin allowing active substances to persist in and/or on the skin to improve absorption and penetration into the epidermis and/or dermis. Similarly, hyaluronic acid can function as a delivery agent for active ingredients into deeper levels of the skin.
The compositions may be delivered to the human subject by applying to the composition topically, in the form of a cream, ointment, gel, or solution. The composition may be applied topically to the skin or mucosa, including skin or mucosa of the face, neck, hands, back and/or décolletage.
The compositions are delivered in an effective amount to improve and/or restore appearance and function to at least one of the skin, mucosa, epidermis, dermis, hypodermis, extracellular matrix, or any combination thereof. The effective amount may be 0.5 mL to 5 mL, wherein any volumes falling therein may serve as endpoints for additional ranges.
In certain aspects, the composition may be administered at predetermined time intervals to achieve desired results. In certain aspects, the compositions are administered daily. In certain aspects, the compositions are administered weekly. In certain aspects, the composition is administered monthly. In certain aspects, the composition is administered bi-weekly. In certain aspects, the composition is administered semi-weekly. In certain aspects, the composition is administered bi-monthly. In certain aspects, the composition is administered semi-monthly.
In certain aspects, the composition may be delivered along with a physical treatment, either before, during or after the physical treatment. The physical treatment may include microneedling, a procedure that includes creating micro-punctures in the skin using miniature sterilized needles. The physical treatment may include dermabrasion, a procedure that removes the top layers of the skin. The physical treatment may include microdermabrasion, a procedure that removes the dead skin from the top layers of the skin. The physical treatment may include chemical peeling, a procedure that includes the use of a, typically acidic, solution to remove the top layers of the skin. The physical treatment may include a laser treatment, such as laser resurfacing—a procedure that utilizes a laser to remove the outer layers of the skin. In certain aspects, the composition is administered concurrently with the physical dermal treatment. In certain aspects, the composition is administered subsequently following the physical dermal treatment. In certain aspects, the composition is administered subsequently before the physical dermal treatment.
In certain aspects, the composition is diluted with an isotonic solution, or amniotic fluid, prior to administration.
In certain aspects, the composition is administered topically. This method includes applying a predetermined amount of the composition to the affected skin or the affected mucosa to improve and/or restore appearance and function to at least one of the skin, mucosa, epidermis, dermis, hypodermis, extracellular matrix, or any combination thereof.
In certain aspects, the composition is administered via intradermal injection. This method includes injecting a predetermined amount of the composition into the dermis of the affected skin or the affected mucosa to improve and/or restore appearance and function to at least one of the skin, mucosa, epidermis, dermis, hypodermis, extracellular matrix, or any combination thereof.
In certain aspects, the composition is administered via subcutaneous injection. This method includes injecting a predetermined amount of the composition into the subcutis below the dermis of the affected skin or the affected mucosa to improve and/or restore appearance and function to at least one of the skin, mucosa, epidermis, dermis, hypodermis, extracellular matrix, or any combination thereof.
In another example, it is further envisioned that the disclosed compositions may have more general applications in the medical field such as general wound packing (occurring in surgical procedures and/or acute trauma resulting in open external and/or internal wounds) and/or wound healing. In this aspect, the aqueous human umbilical cord filtrate may be used alone, or in combination with the micronized human umbilical cord. In some aspects, the micronized human umbilical cord may be combined with amniotic fluid in lieu of aqueous human umbilical cord filtrate. In these aspects, it is envisioned that one would initially assess the wound to generally determine the overall viscosity and thickness of the (mixed) two-part composition needed to, for example, pack and/or treat a subject's wound. Next, one sterilely mixes the composition to an effective viscosity to induce blood clotting; and then sterilely packs the subject's wound with the sterilely mixed composition to induce blood clotting within the sterilely packed wound. In certain aspects, the micronized human umbilical cord composition and the aqueous human umbilical cord filtrate are both sterile and non-immunogenic and are mixed at a ratio of 2:1 to 1:2 micronized human umbilical cord composition and the aqueous human umbilical cord filtrate during this method. If very viscous mixed composition is desired, a higher proportion of the micronized human umbilical cord composition is mixed with a lower proportion the aqueous human umbilical cord filtrate, and conversely, if a less viscous mixed composition is desired, a higher proportion of the aqueous human umbilical cord filtrate is mixed with a lower proportion of the micronized human umbilical cord composition. The above-mentioned packing may be repeated as necessary.
The foregoing description provides embodiments of the invention by way of example only. It is envisioned that other embodiments may perform similar functions and/or achieve similar results. Any and all such equivalent embodiments and examples are within the scope of the present invention and are intended to be covered by the appended claims.
| Number | Date | Country | |
|---|---|---|---|
| 63471592 | Jun 2023 | US |
