US20220098331A1 - Kits and methods of using hyaluronidase to modify polysaccharide fillers and delivery systems - Google Patents
Kits and methods of using hyaluronidase to modify polysaccharide fillers and delivery systems Download PDFInfo
- Publication number
- US20220098331A1 US20220098331A1 US17/548,277 US202117548277A US2022098331A1 US 20220098331 A1 US20220098331 A1 US 20220098331A1 US 202117548277 A US202117548277 A US 202117548277A US 2022098331 A1 US2022098331 A1 US 2022098331A1
- Authority
- US
- United States
- Prior art keywords
- polysaccharide
- agarose
- hyaluronidase
- mass
- drug delivery
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
- 150000004676 glycans Chemical class 0.000 title claims abstract description 64
- 229920001282 polysaccharide Polymers 0.000 title claims abstract description 64
- 239000005017 polysaccharide Substances 0.000 title claims abstract description 64
- 108010003272 Hyaluronate lyase Proteins 0.000 title claims abstract description 50
- 102000001974 Hyaluronidases Human genes 0.000 title claims abstract description 50
- 229960002773 hyaluronidase Drugs 0.000 title claims abstract description 50
- 238000000034 method Methods 0.000 title claims abstract description 13
- 239000000945 filler Substances 0.000 title abstract description 33
- 238000012377 drug delivery Methods 0.000 claims abstract description 20
- 229920000936 Agarose Polymers 0.000 claims description 62
- WQZGKKKJIJFFOK-SVZMEOIVSA-N (+)-Galactose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-SVZMEOIVSA-N 0.000 claims description 20
- 230000002500 effect on skin Effects 0.000 claims description 20
- 235000000346 sugar Nutrition 0.000 claims description 17
- 150000008163 sugars Chemical class 0.000 claims description 17
- 230000000694 effects Effects 0.000 claims description 13
- 229940079593 drug Drugs 0.000 claims description 11
- 239000003814 drug Substances 0.000 claims description 11
- DCQFFOLNJVGHLW-ZNVMLXAYSA-N (1s,4s,5r,8r)-2,6-dioxabicyclo[3.2.1]octane-3,4,8-triol Chemical compound O[C@H]1[C@@]2([H])OC[C@]1([H])OC(O)[C@H]2O DCQFFOLNJVGHLW-ZNVMLXAYSA-N 0.000 claims description 10
- 230000004048 modification Effects 0.000 claims description 8
- 238000012986 modification Methods 0.000 claims description 8
- 241001465754 Metazoa Species 0.000 claims description 7
- 239000012530 fluid Substances 0.000 claims description 5
- 239000003193 general anesthetic agent Substances 0.000 claims description 4
- 239000003795 chemical substances by application Substances 0.000 claims description 2
- 239000000203 mixture Substances 0.000 abstract description 11
- 229940044700 hylenex Drugs 0.000 description 32
- 101710110830 Beta-agarase Proteins 0.000 description 23
- 102000004190 Enzymes Human genes 0.000 description 20
- 108090000790 Enzymes Proteins 0.000 description 20
- 229940088598 enzyme Drugs 0.000 description 20
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 description 17
- 229920002674 hyaluronan Polymers 0.000 description 17
- 229960003160 hyaluronic acid Drugs 0.000 description 17
- 238000002347 injection Methods 0.000 description 13
- 239000007924 injection Substances 0.000 description 13
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 11
- 230000015556 catabolic process Effects 0.000 description 10
- 238000006731 degradation reaction Methods 0.000 description 8
- 238000009472 formulation Methods 0.000 description 8
- XASQSRDYYCWSME-NWCCWNDLSA-N (2S,3R,4S,5R,6R)-4-[[(1S,3S,4S,5S,8R)-8-[(2S,3R,4S,5S,6R)-4-[[(1S,3S,4S,5R,8R)-4,8-dihydroxy-2,6-dioxabicyclo[3.2.1]octan-3-yl]oxy]-3,5-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-4-hydroxy-2,6-dioxabicyclo[3.2.1]octan-3-yl]oxy]-6-(hydroxymethyl)oxane-2,3,5-triol Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O[C@@H]2O[C@H]3CO[C@H]([C@@H]3O[C@@H]3O[C@H](CO)[C@H](O)[C@H](O[C@@H]4O[C@H]5CO[C@H]([C@@H]5O)[C@@H]4O)[C@H]3O)[C@@H]2O)[C@@H]1O XASQSRDYYCWSME-NWCCWNDLSA-N 0.000 description 7
- 239000011543 agarose gel Substances 0.000 description 7
- JWMBOBQNPBCYER-UHFFFAOYSA-N 4-[(4,8-dihydroxy-2,6-dioxabicyclo[3.2.1]octan-3-yl)oxy]-6-(hydroxymethyl)oxane-2,3,5-triol Chemical compound OC1C(CO)OC(O)C(O)C1OC1C(O)C(C2O)OCC2O1 JWMBOBQNPBCYER-UHFFFAOYSA-N 0.000 description 6
- 238000006243 chemical reaction Methods 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 239000000126 substance Substances 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 239000004677 Nylon Substances 0.000 description 5
- 238000003776 cleavage reaction Methods 0.000 description 5
- 239000000470 constituent Substances 0.000 description 5
- 230000007515 enzymatic degradation Effects 0.000 description 5
- 239000012528 membrane Substances 0.000 description 5
- 229920001778 nylon Polymers 0.000 description 5
- 230000007017 scission Effects 0.000 description 5
- 239000006228 supernatant Substances 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 4
- 239000007795 chemical reaction product Substances 0.000 description 4
- 150000002500 ions Chemical class 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- IAJILQKETJEXLJ-UHFFFAOYSA-N Galacturonsaeure Natural products O=CC(O)C(O)C(O)C(O)C(O)=O IAJILQKETJEXLJ-UHFFFAOYSA-N 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 150000002016 disaccharides Chemical group 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 238000001727 in vivo Methods 0.000 description 3
- MSWZFWKMSRAUBD-IVMDWMLBSA-N 2-amino-2-deoxy-D-glucopyranose Chemical compound N[C@H]1C(O)O[C@H](CO)[C@@H](O)[C@@H]1O MSWZFWKMSRAUBD-IVMDWMLBSA-N 0.000 description 2
- 206010003694 Atrophy Diseases 0.000 description 2
- NNJVILVZKWQKPM-UHFFFAOYSA-N Lidocaine Chemical compound CCN(CC)CC(=O)NC1=C(C)C=CC=C1C NNJVILVZKWQKPM-UHFFFAOYSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 238000005903 acid hydrolysis reaction Methods 0.000 description 2
- IAJILQKETJEXLJ-QTBDOELSSA-N aldehydo-D-glucuronic acid Chemical compound O=C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)C(O)=O IAJILQKETJEXLJ-QTBDOELSSA-N 0.000 description 2
- 230000004075 alteration Effects 0.000 description 2
- 230000037444 atrophy Effects 0.000 description 2
- MSWZFWKMSRAUBD-UHFFFAOYSA-N beta-D-galactosamine Natural products NC1C(O)OC(CO)C(O)C1O MSWZFWKMSRAUBD-UHFFFAOYSA-N 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000002537 cosmetic Substances 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- 230000007071 enzymatic hydrolysis Effects 0.000 description 2
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 description 2
- 229960002442 glucosamine Drugs 0.000 description 2
- 229940097043 glucuronic acid Drugs 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 229960004194 lidocaine Drugs 0.000 description 2
- 238000001819 mass spectrum Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 229920001542 oligosaccharide Polymers 0.000 description 2
- 150000002482 oligosaccharides Chemical class 0.000 description 2
- 230000000704 physical effect Effects 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 230000037303 wrinkles Effects 0.000 description 2
- USFZMSVCRYTOJT-UHFFFAOYSA-N Ammonium acetate Chemical compound N.CC(O)=O USFZMSVCRYTOJT-UHFFFAOYSA-N 0.000 description 1
- 239000005695 Ammonium acetate Substances 0.000 description 1
- 101710117542 Botulinum neurotoxin type A Proteins 0.000 description 1
- XQFRJNBWHJMXHO-RRKCRQDMSA-N IDUR Chemical compound C1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C(I)=C1 XQFRJNBWHJMXHO-RRKCRQDMSA-N 0.000 description 1
- OVRNDRQMDRJTHS-UHFFFAOYSA-N N-acelyl-D-glucosamine Natural products CC(=O)NC1C(O)OC(CO)C(O)C1O OVRNDRQMDRJTHS-UHFFFAOYSA-N 0.000 description 1
- OVRNDRQMDRJTHS-FMDGEEDCSA-N N-acetyl-beta-D-glucosamine Chemical group CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O OVRNDRQMDRJTHS-FMDGEEDCSA-N 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 108010045649 agarase Proteins 0.000 description 1
- AEMOLEFTQBMNLQ-WAXACMCWSA-N alpha-D-glucuronic acid Chemical group O[C@H]1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O AEMOLEFTQBMNLQ-WAXACMCWSA-N 0.000 description 1
- 229940043376 ammonium acetate Drugs 0.000 description 1
- 235000019257 ammonium acetate Nutrition 0.000 description 1
- 229940089093 botox Drugs 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000007857 degradation product Substances 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 239000000539 dimer Substances 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 238000005243 fluidization Methods 0.000 description 1
- 238000013467 fragmentation Methods 0.000 description 1
- 238000006062 fragmentation reaction Methods 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 230000036571 hydration Effects 0.000 description 1
- 238000006703 hydration reaction Methods 0.000 description 1
- 229960004716 idoxuridine Drugs 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000013580 millipore water Substances 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 229950006780 n-acetylglucosamine Drugs 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000004451 qualitative analysis Methods 0.000 description 1
- 230000036632 reaction speed Effects 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000005464 sample preparation method Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
- C08B37/0036—Galactans; Derivatives thereof
- C08B37/0039—Agar; Agarose, i.e. D-galactose, 3,6-anhydro-D-galactose, methylated, sulfated, e.g. from the red algae Gelidium and Gracilaria; Agaropectin; Derivatives thereof, e.g. Sepharose, i.e. crosslinked agarose
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
- A61K8/66—Enzymes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
- A61K8/73—Polysaccharides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
- A61K8/73—Polysaccharides
- A61K8/735—Mucopolysaccharides, e.g. hyaluronic acid; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/20—Polysaccharides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/54—Biologically active materials, e.g. therapeutic substances
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/06—Ointments; Bases therefor; Other semi-solid forms, e.g. creams, sticks, gels
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
- A61L2300/252—Polypeptides, proteins, e.g. glycoproteins, lipoproteins, cytokines
- A61L2300/254—Enzymes, proenzymes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2400/00—Materials characterised by their function or physical properties
- A61L2400/06—Flowable or injectable implant compositions
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/34—Materials or treatment for tissue regeneration for soft tissue reconstruction
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01035—Hyaluronoglucosaminidase (3.2.1.35), i.e. hyaluronidase
Definitions
- Embodiments of the present invention were not conceived or reduced to practice with Federal sponsorship.
- Polysaccharides are used as cosmetic fillers and as a component of injectable drug delivery systems. When polysaccharides are placed in the body as a filler or as a drug delivery system, the polysaccharides may have a long period of time before such are resorbed into the body through metabolism or other means.
- Polysaccharides used as fillers have certain properties such as viscosity, resistance to degradation, texture, feel, resistance to pressure and the like. These properties are dictated by the organisms from which the polysaccharides are derived or isolated from.
- Embodiments of the present invention facilitate the use of polysaccharides, particularly as such polysaccharides are used as fillers and drug delivery vehicles.
- Embodiments of the present invention allow the polysaccharide to be modified in vivo or in vitro to impart special physical and chemical properties. For example, without limitation, the physical properties of texture, feel, resistance to pressure, viscosity and the like can be altered or modified, in vivo or in vitro. Similarly, the chemical properties, resistance to degradation and speed, of degradation can be modified or altered.
- polysaccharide fillers used for cosmetic purposes typically comprise solutions of 1.0 to 5.0%. These solutions may also comprise other materials to improve the flow or feel of the filler, such as hyaluronic acid, and anesthetic agents to address the potential discomfort during the administration of the filler. Anesthetic agents are well known in the art and include, without limitation, lidocaine. These fillers are administered to the deep layer of the skin. An example of a filler of this type is described in PCT/IB2014/060322 to Ghimas SPA, the entire content of which is incorporated herein by reference.
- One embodiment of the present invention is directed to a method of altering or modifying a mass comprising a polysaccharide held in the body of an animal.
- the method comprises the steps of administering an effective amount of hyaluronidase to the mass.
- the hyaluronidase can be administered to the mass prior to the mass being placed in the body or after the mass has been placed in the body.
- One embodiment of the method features a polysaccharide having one or more sugars selected from the group consisting of D-galactose and 3, 6-anhydro-L-galactopyranose. These sugars are the constituents of agarose. Agarose is used as a dermal filler and as a drug delivery vehicle. As used herein, a “drug delivery vehicle” comprises a polysaccharide and one or more drugs.
- hyaluronidase refers to an enzyme which cleave Beta-1,4 bond of hyaluronic acid.
- the term hyaluronidase includes its salts and derivatives which retain its enzymatic activity.
- a recombinant form of hyaluronidase for human injection sold under the trademark Hylenex® (Halozyme, Inc., San Diego, Calif.), is an FDA approved enzyme, and the current standard for the off-label use of treating overcorrection of hyaluronic acid (HA) based dermal fillers.
- Hyaluronidase hydrolyzes HA by cleaving the ⁇ -1,4 bond between the glucosamine and glucuronic acid.
- hyaluronidase alters the form of the polysaccharide, increasing its dissolution, breaking down and fluidizing the mass. It is surprising and unexpected because enzymes such as hyaluronidase are highly specific in the substrate.
- the polysaccharide having one or more sugars selected from the group consisting of D-galactose and 3, 6-anhydro-L-galactopyranose, such as agarose is one such polysaccharide to which hyaluronidase acts upon.
- an effective amount refers to an amount to cause the polysaccharide to assume a desired fluidized form.
- the desired fluidized form refers to a more fluid form compared to agarose of same polymer composition and hydration without a hyaluronidase being present.
- the hyaluronidase has utility to facilitate removal of the polysaccharide, control the delivery or release of drug, address hardness, graininess or nodules in the mass and minimize the mark on the skin from the injection of drug.
- one embodiment of an invention directed to a drug delivery system comprises a polysaccharide and hyaluronidase held in a vessel for reconstitution. Upon reconstitution, the hyaluronidase is injected into a mass of the polysaccharide to act on the polysaccharide and render the polysaccharide in a more fluid state.
- drug is used to refer to any compound or compounds used to effect a biological change or treat a medical condition.
- the drug may be incorporated into the polysaccharide prior to reconstitution or after reconstitution.
- examples of drugs include without limitation, lidocaine, and other anesthetic agents, onabotulinemtoxin A (BOTOX®, Allergan) and other aesthetic agents.
- a further embodiment of the present invention is directed to a kit for performing dermal filling procedures or for the administration of drug.
- the kit comprises a polysaccharide for forming a mass in the body of an animal and a hyaluronidase for maintaining or making the polysaccharide more fluid for administration to the mass held in the body to effect a modification, correction or speeding the resorption of the mass.
- One embodiment of the present kit features a polysaccharide having one or more sugars selected from the group consisting of D-galactose and 3, 6-anhydro-L-galactopyranose. These sugars are constituents of the polysaccharide, agarose.
- a further embodiment of the present invention is directed to a formulation for a dermal filler.
- the formulation comprises the reaction product of agarose and hyaluronidase.
- One reaction product is neoagarohexaose.
- One formulation comprises agarose and neoagarohexarose. Neoagarohexarose is added to agarose to create a dermal filler or drug delivery vehicle with properties of desire texture, viscosity, injectability and resistance to degradation.
- FIG. 1 depicts a kit embodying features of the present invention
- FIG. 2A shows the structure of agarose
- FIG. 2B illustrates cleavages on agarose, having features of the present invention
- FIG. 3 is an image showing a cleavage on HA
- FIG. 4 is an ion chromatogram of a sample, having features of the present invention.
- FIG. 5 is another ion chromatogram of a sample, having features of the present invention.
- Embodiments of the present invention will now be described in detail with respect to a kit for performing dermal filling procedures or for the administration of drug.
- the present description is directed to embodiments which are considered to be the best mode to practice the present invention at the time of the writing of the present description.
- Those skilled in the art will recognize that the understanding of the best mode may change in time.
- Those skilled in the art will also recognize that the features of the present invention described are subject to alteration and modification and such that the present discussion should not be considered limiting.
- kit 11 a kit, generally designated by the numeral 11 , embodying features of the present invention is depicted.
- the term “kit” is used to denote a bundled assembly of parts and constituents for performing a method.
- the kit 11 comprises two vials, a first vial 13 containing a polysaccharide for forming a mass in the body of an animal and a second vial 15 containing hyaluronidase for fluidizing the polysaccharide for administration to the mass held in the body.
- the first vial 13 and second vial 15 are bundled or held in a package 17 with means for administering the polysaccharide and means for administering the enzyme, in the form of first syringe 19 and second syringe 21 , and instructions 23 .
- the first vial 13 contains a polysaccharide for forming a mass in the body of an animal or patient.
- the polysaccharide can be pre-made and in a final form ready for administration or be lyophilized for reconstitution with water.
- the first syringe 19 is used to withdraw the reconstituted or pre-made polysaccharide for injection into an animal or patient.
- a human subject may use the mass to conceal wrinkles or to build bulk to areas of the body showing signs of atrophy or for which a fuller appearance is desired.
- the present kit features a polysaccharide having one or more sugars selected from the group consisting of D-galactose and 3, 6-anhydro-L-galactopyranose. These sugars are constituents of the polysaccharide, agarose, which is used as a dermal filler. Agarose is sold as a dermal filler under the mark ALGINESS®.
- the second vial 15 contains hyaluronidase to effect a modification or speeding the resorption or altering the fluidity of the mass.
- the hyaluronidase can be pre-made and in a final form ready for administration or be lyophilized for reconstitution with water.
- the second syringe 21 is used to withdraw the reconstituted or pre-made hyaluronidase for injection into the mass.
- the subject may desire the mass to have a softer feel, or the mass may exhibit graininess or nodules or the mass may have been overinjected.
- the term, “overinjected” refers to a condition in which the mass appears too large for the desired effect.
- the hyaluronidase is placed in the mass and the mass manipulated to distribute the hyaluronidase where the hyaluronidase may effect a modification of the polysaccharide.
- the polysaccharide assumes a more fluid form, capable of faster metabolization, removing graininess and nodules and presenting a softer mass and/or a mass which is reduced or will be reduced in size more quickly.
- Embodiments of the present invention facilitate the use of polysaccharides, particularly as such polysaccharides are used as fillers and drug delivery vehicles.
- Embodiments of the present invention allow the polysaccharide to be modified in vivo or in vitro to impart special physical and chemical properties. That is, the hyaluronidase can be distributed into the polysaccharide prior to administration of the polysaccharide or after administration of the polysaccharide to create a desired physical property of texture, feel, resistance to pressure, viscosity and the like. Similarly, the chemical properties of resistance to degradation and speed of degradation can be modified or altered.
- the modification of physical and chemical features of a polysaccharide used as a filler or drug delivery system allows the health practitioner to modify the filler mass after it has been placed in the body or prior to placement in the body.
- the mass may be fluidized for removal or minimization by withdrawing the mass after the administration of the hyaluronidase.
- the fluidization of the mass allows the mass to be redistributed and aids in the resorption of the mass.
- kit 11 and the instructions 23 will now be described with respect to an embodiment of the present invention directed to a method of altering or modifying a mass comprising a polysaccharide held in the body of an animal.
- the instructions 23 direct the user to reconstitute the polysaccharide held in first vial 13 and the hyaluronidase held in second vial 15 if reconstitution is needed.
- the instructions 23 direct the user to administer polysaccharide held in the first vial 13 .
- the instructions 23 further direct the user to administer an effective amount of hyaluronidase for the polysaccharide to the mass.
- the hyaluronidase can be administered to the mass prior to the mass being placed in the body or after the mass has been placed in the body.
- More than one application of hyaluronidase may be used to obtain the desired consistency of the mass.
- the mass may be gently kneaded or manipulated to distribute the hyaluronidase throughout the mass structure.
- the fluidized mass may be manipulated into a desired position or removed by suction through the same syringe administering the hyaluronidase.
- Embodiments of the method and kits feature a polysaccharide having one or more sugars selected from the group consisting of D-galactose and 3, 6-anhydro-L-galactopyranose. These sugars are the constituents of agarose. Agarose is used as a dermal filler and as a drug delivery vehicle.
- Embodiments of the method and kits features hyaluronidase.
- Hyaluronidase is well tolerated and the amount is based on the mass of polysaccharide. Monographs, for such enzyme, are available from the respective manufacturers and are incorporated herein by reference.
- a further embodiment of the present invention is directed to a formulation for a dermal filler.
- the formulation comprises the reaction product of agarose and hyaluronidase with agarose.
- One reaction product is neoagarohexaose.
- One formulation comprises agarose and neoagarohexarose. Neoagarohexarose is added to agarose to create a dermal filler or drug delivery vehicle with properties of desire texture, viscosity, injectability and resistance to degradation.
- the relative percentages of agarose and neoagarosehexaose are chosen for the desired texture, viscosity, injectability and resistance to degradation.
- formulations with higher percentages of neoagarosehexaose and lower percentages of agarose may show higher rates of resorption in the body, greater injectability, lower viscosity and smoother texture.
- One embodiment of the present formulation comprises a range of 99-50% agarose with the remainder neoagarosehexaose, another embodiment features 95-75% agarose with the remainder neoagarosehexaose, and another embodiment features 99-90% agarose and the remainder neoagarosehexaose.
- Hylenex® recombinant (hyaluronidase human injection) is the so-called gold standard for treating over-injection or negative side-effects of hyaluronic acid dermal fillers.
- Hyaluronidase cleaves the ⁇ -1,4 linkage of hyaluronic acid. While the structure of hyaluronic acid and agarose are very different, it is possible for hyaluronidase to hydrolyze agarose at the ⁇ -1,4 bond.
- ⁇ -agarase cleaved agarose at the ⁇ -1,4 glyosidic linkages producing the dimer, tetramer and hexamer neoagaro oligosaccharides. The presence of the hexamer was also observed in the agarose sample exposed to Hylenex®.
- agarose is linear polymer composed of repeating units of D-galactose and 3,6-anhydro-L-galactopyranose, linked by ⁇ -1,3 and ⁇ -1,4 glycosidic bonds.
- FIG. 2B shows that ⁇ -agarase hydrolyzes by cleaving agarose at the ⁇ -1,4 bond between D-galactose (G) and 3,6-anhydro-L-galactopyranose (A), producing a series of neoagarooligosaccharides with repeating disaccharide units.
- neoagarooligosaccharides of varying molecular mass can be produced, including neoagarobiose (324.28 g/mol), neoagarotetraose (630.55 g/mol) and neoagarohexaose (936.82 g/mol).
- Hylenex® recombinant (hyaluronidase human injection) is an FDA approved enzyme, and current standard for the off-label use of treating overcorrection of hyaluronic acid (HA) based dermal fillers.
- HA hyaluronic acid
- 2-4 HA is composed of repeating units of N-Acetyl-D-Glucosamine and D-Glucuronic acid, linked by ⁇ -1,3 and ( ⁇ -1,4 bonds.
- Hyaluronidase hydrolyzes HA by cleaving the ⁇ -1,4 bond between the glucosamine and glucuronic acid, producing a series of repeating disaccharide units, as shown in FIG. 3 (Image from: www.sigmaaldrich.com).
- the objective of this study was to determine if Hylenex® recombinant hydrolyzes the agarose-based dermal filler, Algeness® DF, based on the cleavage of the ⁇ -1,4 linkage D-galactose and 3,6-anhydro-L-galactopyranose.
- Neoagarotetraose was used as a reference standard to confirm enzymatic activity in this study.
- Agarose powder (0.2 g) was added to 100 ml of water (99° C.) and cooled in a water bath to 42° C.
- Three control samples were prepared using 1000 ⁇ l of molten agarose (2 mg). Samples were incubated at 42° C. for 2 hours, and then either 1) centrifuged at 3000 g for 5 min at 4° C., 2) centrifuged at 12000 g for 5 min at 4° C. or 3) heated at 90° C. for 10 minutes (“kill enzyme” step) and centrifuged at 12000 g for 5 min at 4° C. Supernatant was filtered using 0.2 ⁇ m Nylon membrane filters. All samples were analyzed by LC/MS to determine if sample preparation methods degraded the agarose gel. Additionally, the Molisch test for sugars was conducted on all samples.
- Hylenex® recombinant hyaluronidase human injection
- ⁇ -agarase ⁇ -agarase to compare the effects of enzymatic degradation on the two samples. Briefly, Hylenex® (200 ⁇ l-30 units) was added to agarose (200 ⁇ l-0.4 mg agarose) and incubated at 37° C. for 2 hours. Additionally, ⁇ -agarase (5 ⁇ l-5 units) was added to agarose (1000 ⁇ l-2 mg agarose) and incubated at 37° C. for 2 hours. Samples were centrifuged at 12000 g for 5 min at 4° C. The supernatant was filtered using 0.2 ⁇ m Nylon membrane filters and analyzed by LC/MS. Additionally, the Molisch test for sugars was conducted on all samples.
- Algeness® DF (3.5% agarose) and Juvederm® Ultra Plus XC (2.4% HA) were exposed to Hylenex® recombinant (hyaluronidase human injection) to compare the effects of enzymatic degradation.
- Algeness® DF was exposed to ⁇ -agarase for comparison. Briefly, enzyme was added to the dermal filler and stirred (800 rpm) at room temperature for 6 hours. Table 1 shows the amount of dermal filler and enzyme used for each reaction. After the reaction, samples were heated at 90° C. for 10 minutes (“kill enzyme”) and then centrifuged at 10000 g for 5 min at 4° C. Supernatant was filtered using 0.2 ⁇ m Nylon membrane filters and analyzed by LC/MS. Additionally, the Molisch test for sugars was conducted on all samples.
- the MS was operated in positive ionization mode for agarose and negative ionization mode for hyaluronic acid, with a fragmentation voltage of 100V. Spectra were recorded in full scan mode from 300 to 2000 m/z. Neoagarotetraose, a known degradation product of agarose when exposed to ⁇ -agarase, was used to confirm enzyme activity.
- Hylenex® recombinant hyaluronidase human injection
- ⁇ -agarase ⁇ -agarase to compare the effects of enzymatic degradation. Briefly, Hylenex® (200 ⁇ l-30 units) was added to agarose (400 ⁇ l-14 mg agarose) and incubated at 37° C. for 2 hours. Additionally, ⁇ -agarase (30 ⁇ l-30 units) was added to agarose (400 ⁇ l-14 mg agarose) and incubated at 37° C. for 2 hours. Samples were centrifuged at 10000 g for 5 min at 4° C. Supernatant was filtered using 0.2 ⁇ m Nylon membrane filters and analyzed by LC/MS. Additionally, the Molisch test was conducted on all samples.
- Algeness® DF (3.5% agarose) and Juvederm® Ultra Plus XC (2.4% HA) were exposed to Hylenex® recombinant (hyaluronidase human injection) to compare the effects of enzymatic degradation.
- Algeness® DF was also exposed to ⁇ -agarase for comparison. Briefly, enzyme was added to the dermal filler and incubated at 37° C. for 48 hours. Table 3 shows the amount of dermal filler and enzyme used for each reaction. After incubation period, samples were heated at 90° C. for 10 minutes (“kill enzyme”) and then centrifuged at 10000 g for 5 min at 4° C. Supernatant was filtered using 0.2 ⁇ m Nylon membrane filters and analyzed by LC/MS. Additionally, the Molisch test for sugars was conducted on all samples.
- FIG. 4 includes two sub-figures, the top one is an ion chromatogram for the three products represented by three peaks designated 1; 2 and 3, and the bottom one consists of three mass spectra, corresponding respectively to the three products.
- LC/MS analysis of the agarose/Hylenex® sample revealed the hexasaccharide only, as shown in FIG. 5 .
- FIG. 5 also includes two sub-figures, the top one is an ion chromatogram for the hexasaccharide represented by a chromatographic peak, and the bottom one is a mass spectrum corresponding to the hexasaccharide.
- the Molisch test is a qualitative analysis used to indicate the presence of carbohydrates in a sample. A positive result is indicated by a purple ring between the sample and the concentrated acid. If sugars are not present in the sample, the solution will remain clear. The presence of monosaccharides results in a faster reaction, while disaccharides and polysaccharides will result in a slower reaction time.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Chemical & Material Sciences (AREA)
- Epidemiology (AREA)
- Medicinal Chemistry (AREA)
- Birds (AREA)
- Dermatology (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Gerontology & Geriatric Medicine (AREA)
- Transplantation (AREA)
- Oral & Maxillofacial Surgery (AREA)
- Materials Engineering (AREA)
- Organic Chemistry (AREA)
- Polymers & Plastics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biochemistry (AREA)
- Inorganic Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Biomedical Technology (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicinal Preparation (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
Embodiments of the present invention are directed to kits, compositions and methods for modifying and altering polysaccharide fillers and drug delivery systems with the application of hyaluronidase
Description
- This application claims priority to U.S. Provisional Application Ser. No. 62/460,756, filed on Feb. 18, 2017, the contents of which are incorporated herein by reference.
- Embodiments of the present invention were not conceived or reduced to practice with Federal sponsorship.
- Polysaccharides are used as cosmetic fillers and as a component of injectable drug delivery systems. When polysaccharides are placed in the body as a filler or as a drug delivery system, the polysaccharides may have a long period of time before such are resorbed into the body through metabolism or other means.
- Polysaccharides used as fillers have certain properties such as viscosity, resistance to degradation, texture, feel, resistance to pressure and the like. These properties are dictated by the organisms from which the polysaccharides are derived or isolated from.
- Embodiments of the present invention facilitate the use of polysaccharides, particularly as such polysaccharides are used as fillers and drug delivery vehicles. Embodiments of the present invention allow the polysaccharide to be modified in vivo or in vitro to impart special physical and chemical properties. For example, without limitation, the physical properties of texture, feel, resistance to pressure, viscosity and the like can be altered or modified, in vivo or in vitro. Similarly, the chemical properties, resistance to degradation and speed, of degradation can be modified or altered.
- The modification of physical and chemical features of a polysaccharide used as a filler or drug delivery system allows the health practitioner to modify the filler mass after it has been placed in the body or prior to placement in the body. As used herein, the term, “mass” refers to the polysaccharide material and the surrounding space it occupies. For example, polysaccharide fillers used for cosmetic purposes typically comprise solutions of 1.0 to 5.0%. These solutions may also comprise other materials to improve the flow or feel of the filler, such as hyaluronic acid, and anesthetic agents to address the potential discomfort during the administration of the filler. Anesthetic agents are well known in the art and include, without limitation, lidocaine. These fillers are administered to the deep layer of the skin. An example of a filler of this type is described in PCT/IB2014/060322 to Ghimas SPA, the entire content of which is incorporated herein by reference.
- One embodiment of the present invention is directed to a method of altering or modifying a mass comprising a polysaccharide held in the body of an animal. The method comprises the steps of administering an effective amount of hyaluronidase to the mass. The hyaluronidase can be administered to the mass prior to the mass being placed in the body or after the mass has been placed in the body.
- One embodiment of the method features a polysaccharide having one or more sugars selected from the group consisting of D-galactose and 3, 6-anhydro-L-galactopyranose. These sugars are the constituents of agarose. Agarose is used as a dermal filler and as a drug delivery vehicle. As used herein, a “drug delivery vehicle” comprises a polysaccharide and one or more drugs.
- As used herein, the term “hyaluronidase” refers to an enzyme which cleave Beta-1,4 bond of hyaluronic acid. The term hyaluronidase includes its salts and derivatives which retain its enzymatic activity. A recombinant form of hyaluronidase for human injection, sold under the trademark Hylenex® (Halozyme, Inc., San Diego, Calif.), is an FDA approved enzyme, and the current standard for the off-label use of treating overcorrection of hyaluronic acid (HA) based dermal fillers. Hyaluronidase hydrolyzes HA by cleaving the β-1,4 bond between the glucosamine and glucuronic acid.
- Surprising and unexpectedly, hyaluronidase alters the form of the polysaccharide, increasing its dissolution, breaking down and fluidizing the mass. It is surprising and unexpected because enzymes such as hyaluronidase are highly specific in the substrate. The polysaccharide having one or more sugars selected from the group consisting of D-galactose and 3, 6-anhydro-L-galactopyranose, such as agarose is one such polysaccharide to which hyaluronidase acts upon. As used herein, the term “an effective amount” refers to an amount to cause the polysaccharide to assume a desired fluidized form. The desired fluidized form refers to a more fluid form compared to agarose of same polymer composition and hydration without a hyaluronidase being present.
- Wherein the mass is a delivery vehicle for one or more drugs, the hyaluronidase has utility to facilitate removal of the polysaccharide, control the delivery or release of drug, address hardness, graininess or nodules in the mass and minimize the mark on the skin from the injection of drug. For example, without limitation, one embodiment of an invention directed to a drug delivery system comprises a polysaccharide and hyaluronidase held in a vessel for reconstitution. Upon reconstitution, the hyaluronidase is injected into a mass of the polysaccharide to act on the polysaccharide and render the polysaccharide in a more fluid state.
- As used herein, the term “drug” is used to refer to any compound or compounds used to effect a biological change or treat a medical condition. The drug may be incorporated into the polysaccharide prior to reconstitution or after reconstitution. Examples of drugs include without limitation, lidocaine, and other anesthetic agents, onabotulinemtoxin A (BOTOX®, Allergan) and other aesthetic agents.
- A further embodiment of the present invention is directed to a kit for performing dermal filling procedures or for the administration of drug. The kit comprises a polysaccharide for forming a mass in the body of an animal and a hyaluronidase for maintaining or making the polysaccharide more fluid for administration to the mass held in the body to effect a modification, correction or speeding the resorption of the mass.
- One embodiment of the present kit features a polysaccharide having one or more sugars selected from the group consisting of D-galactose and 3, 6-anhydro-L-galactopyranose. These sugars are constituents of the polysaccharide, agarose.
- A further embodiment of the present invention is directed to a formulation for a dermal filler. The formulation comprises the reaction product of agarose and hyaluronidase. One reaction product is neoagarohexaose. One formulation comprises agarose and neoagarohexarose. Neoagarohexarose is added to agarose to create a dermal filler or drug delivery vehicle with properties of desire texture, viscosity, injectability and resistance to degradation.
- These and other features and advantages will be apparent to those skilled in the art upon viewing the figure which is briefly described below and studying the details description that follows.
-
FIG. 1 depicts a kit embodying features of the present invention; -
FIG. 2A shows the structure of agarose; -
FIG. 2B illustrates cleavages on agarose, having features of the present invention; -
FIG. 3 is an image showing a cleavage on HA; -
FIG. 4 is an ion chromatogram of a sample, having features of the present invention; and -
FIG. 5 is another ion chromatogram of a sample, having features of the present invention. - Embodiments of the present invention will now be described in detail with respect to a kit for performing dermal filling procedures or for the administration of drug. The present description is directed to embodiments which are considered to be the best mode to practice the present invention at the time of the writing of the present description. Those skilled in the art will recognize that the understanding of the best mode may change in time. Those skilled in the art will also recognize that the features of the present invention described are subject to alteration and modification and such that the present discussion should not be considered limiting.
- Turning now to
FIG. 1 , a kit, generally designated by the numeral 11, embodying features of the present invention is depicted. The term “kit” is used to denote a bundled assembly of parts and constituents for performing a method. Thekit 11 comprises two vials, a first vial 13 containing a polysaccharide for forming a mass in the body of an animal and a second vial 15 containing hyaluronidase for fluidizing the polysaccharide for administration to the mass held in the body. The first vial 13 and second vial 15 are bundled or held in a package 17 with means for administering the polysaccharide and means for administering the enzyme, in the form offirst syringe 19 andsecond syringe 21, andinstructions 23. - The first vial 13 contains a polysaccharide for forming a mass in the body of an animal or patient. The polysaccharide can be pre-made and in a final form ready for administration or be lyophilized for reconstitution with water. The
first syringe 19 is used to withdraw the reconstituted or pre-made polysaccharide for injection into an animal or patient. For example, a human subject may use the mass to conceal wrinkles or to build bulk to areas of the body showing signs of atrophy or for which a fuller appearance is desired. - The present kit features a polysaccharide having one or more sugars selected from the group consisting of D-galactose and 3, 6-anhydro-L-galactopyranose. These sugars are constituents of the polysaccharide, agarose, which is used as a dermal filler. Agarose is sold as a dermal filler under the mark ALGINESS®.
- The second vial 15 contains hyaluronidase to effect a modification or speeding the resorption or altering the fluidity of the mass. The hyaluronidase can be pre-made and in a final form ready for administration or be lyophilized for reconstitution with water. The
second syringe 21 is used to withdraw the reconstituted or pre-made hyaluronidase for injection into the mass. For example, in a human subject using the mass to conceal wrinkles or to build bulk to areas of the body showing signs of atrophy, the subject may desire the mass to have a softer feel, or the mass may exhibit graininess or nodules or the mass may have been overinjected. As used herein the term, “overinjected” refers to a condition in which the mass appears too large for the desired effect. The hyaluronidase is placed in the mass and the mass manipulated to distribute the hyaluronidase where the hyaluronidase may effect a modification of the polysaccharide. The polysaccharide assumes a more fluid form, capable of faster metabolization, removing graininess and nodules and presenting a softer mass and/or a mass which is reduced or will be reduced in size more quickly. - Embodiments of the present invention facilitate the use of polysaccharides, particularly as such polysaccharides are used as fillers and drug delivery vehicles. Embodiments of the present invention allow the polysaccharide to be modified in vivo or in vitro to impart special physical and chemical properties. That is, the hyaluronidase can be distributed into the polysaccharide prior to administration of the polysaccharide or after administration of the polysaccharide to create a desired physical property of texture, feel, resistance to pressure, viscosity and the like. Similarly, the chemical properties of resistance to degradation and speed of degradation can be modified or altered.
- The modification of physical and chemical features of a polysaccharide used as a filler or drug delivery system allows the health practitioner to modify the filler mass after it has been placed in the body or prior to placement in the body. For example, the mass may be fluidized for removal or minimization by withdrawing the mass after the administration of the hyaluronidase. The fluidization of the mass allows the mass to be redistributed and aids in the resorption of the mass.
- The use of the
kit 11 and theinstructions 23 will now be described with respect to an embodiment of the present invention directed to a method of altering or modifying a mass comprising a polysaccharide held in the body of an animal. Theinstructions 23 direct the user to reconstitute the polysaccharide held in first vial 13 and the hyaluronidase held in second vial 15 if reconstitution is needed. Theinstructions 23 direct the user to administer polysaccharide held in the first vial 13. Theinstructions 23 further direct the user to administer an effective amount of hyaluronidase for the polysaccharide to the mass. The hyaluronidase can be administered to the mass prior to the mass being placed in the body or after the mass has been placed in the body. More than one application of hyaluronidase may be used to obtain the desired consistency of the mass. The mass may be gently kneaded or manipulated to distribute the hyaluronidase throughout the mass structure. The fluidized mass may be manipulated into a desired position or removed by suction through the same syringe administering the hyaluronidase. - Embodiments of the method and kits feature a polysaccharide having one or more sugars selected from the group consisting of D-galactose and 3, 6-anhydro-L-galactopyranose. These sugars are the constituents of agarose. Agarose is used as a dermal filler and as a drug delivery vehicle.
- Embodiments of the method and kits features hyaluronidase. Hyaluronidase is well tolerated and the amount is based on the mass of polysaccharide. Monographs, for such enzyme, are available from the respective manufacturers and are incorporated herein by reference.
- A further embodiment of the present invention is directed to a formulation for a dermal filler. The formulation comprises the reaction product of agarose and hyaluronidase with agarose. One reaction product is neoagarohexaose. One formulation comprises agarose and neoagarohexarose. Neoagarohexarose is added to agarose to create a dermal filler or drug delivery vehicle with properties of desire texture, viscosity, injectability and resistance to degradation.
- The relative percentages of agarose and neoagarosehexaose are chosen for the desired texture, viscosity, injectability and resistance to degradation. For example, without limitation, formulations with higher percentages of neoagarosehexaose and lower percentages of agarose may show higher rates of resorption in the body, greater injectability, lower viscosity and smoother texture. One embodiment of the present formulation comprises a range of 99-50% agarose with the remainder neoagarosehexaose, another embodiment features 95-75% agarose with the remainder neoagarosehexaose, and another embodiment features 99-90% agarose and the remainder neoagarosehexaose.
- These and other features will be apparent form the following Example.
- Methods: First, pure agarose was exposed to Hylenex® recombinant (hyaluronidase human injection) and β-agarase. Liquid chromatography/mass spectrometry (LC/MS) was used to detect the presence of oligosaccharides which are known breakdown products of agarose. Finally, the effects of hyaluronidase and β-agarase were compared on Algeness® dermal fillers.
- Results: Hylenex® recombinant (hyaluronidase human injection) is the so-called gold standard for treating over-injection or negative side-effects of hyaluronic acid dermal fillers. Hyaluronidase cleaves the β-1,4 linkage of hyaluronic acid. While the structure of hyaluronic acid and agarose are very different, it is possible for hyaluronidase to hydrolyze agarose at the β-1,4 bond. As expected, β-agarase cleaved agarose at the β-1,4 glyosidic linkages producing the dimer, tetramer and hexamer neoagaro oligosaccharides. The presence of the hexamer was also observed in the agarose sample exposed to Hylenex®.
- Conclusions: The enzymatic hydrolysis of pure agarose by β-agarase produced three compounds, neoagarobiose, neoagarotetraose and neoagarohexaose, which result from the cleavage of β-1,4 glycosidic bonds. The enzymatic hydrolysis of agarose by Hylenex® produced one product, neoagarohexaose. Similar results were observed in the enzymatic degradation of Algeness® DF when exposed to β-agarase and Hylenex®.
- As shown in
FIG. 2A , agarose is linear polymer composed of repeating units of D-galactose and 3,6-anhydro-L-galactopyranose, linked by α-1,3 and β-1,4 glycosidic bonds.FIG. 2B shows that β-agarase hydrolyzes by cleaving agarose at the β-1,4 bond between D-galactose (G) and 3,6-anhydro-L-galactopyranose (A), producing a series of neoagarooligosaccharides with repeating disaccharide units.1 Depending on the cleavage site, neoagarooligosaccharides of varying molecular mass can be produced, including neoagarobiose (324.28 g/mol), neoagarotetraose (630.55 g/mol) and neoagarohexaose (936.82 g/mol). - Hylenex® recombinant (hyaluronidase human injection) is an FDA approved enzyme, and current standard for the off-label use of treating overcorrection of hyaluronic acid (HA) based dermal fillers.2-4 HA is composed of repeating units of N-Acetyl-D-Glucosamine and D-Glucuronic acid, linked by β-1,3 and (β-1,4 bonds. Hyaluronidase hydrolyzes HA by cleaving the β-1,4 bond between the glucosamine and glucuronic acid, producing a series of repeating disaccharide units, as shown in
FIG. 3 (Image from: www.sigmaaldrich.com). - The objective of this study was to determine if Hylenex® recombinant hydrolyzes the agarose-based dermal filler, Algeness® DF, based on the cleavage of the β-1,4 linkage D-galactose and 3,6-anhydro-L-galactopyranose. Neoagarotetraose was used as a reference standard to confirm enzymatic activity in this study.
- Control Samples—0.2% Agarose Gel
- Agarose powder (0.2 g) was added to 100 ml of water (99° C.) and cooled in a water bath to 42° C. Three control samples were prepared using 1000 μl of molten agarose (2 mg). Samples were incubated at 42° C. for 2 hours, and then either 1) centrifuged at 3000 g for 5 min at 4° C., 2) centrifuged at 12000 g for 5 min at 4° C. or 3) heated at 90° C. for 10 minutes (“kill enzyme” step) and centrifuged at 12000 g for 5 min at 4° C. Supernatant was filtered using 0.2 μm Nylon membrane filters. All samples were analyzed by LC/MS to determine if sample preparation methods degraded the agarose gel. Additionally, the Molisch test for sugars was conducted on all samples.
- Enzyme Degradation—0.2% Agarose Gel
- Agarose gel (0.2%) was exposed to Hylenex® recombinant (hyaluronidase human injection) and β-agarase to compare the effects of enzymatic degradation on the two samples. Briefly, Hylenex® (200 μl-30 units) was added to agarose (200 μl-0.4 mg agarose) and incubated at 37° C. for 2 hours. Additionally, β-agarase (5 μl-5 units) was added to agarose (1000 μl-2 mg agarose) and incubated at 37° C. for 2 hours. Samples were centrifuged at 12000 g for 5 min at 4° C. The supernatant was filtered using 0.2 μm Nylon membrane filters and analyzed by LC/MS. Additionally, the Molisch test for sugars was conducted on all samples.
- Enzyme Degradation—Algeness® DF and Juvederm® Ultra Plus XC
- Algeness® DF (3.5% agarose) and Juvederm® Ultra Plus XC (2.4% HA) were exposed to Hylenex® recombinant (hyaluronidase human injection) to compare the effects of enzymatic degradation. In addition, Algeness® DF was exposed to β-agarase for comparison. Briefly, enzyme was added to the dermal filler and stirred (800 rpm) at room temperature for 6 hours. Table 1 shows the amount of dermal filler and enzyme used for each reaction. After the reaction, samples were heated at 90° C. for 10 minutes (“kill enzyme”) and then centrifuged at 10000 g for 5 min at 4° C. Supernatant was filtered using 0.2 μm Nylon membrane filters and analyzed by LC/MS. Additionally, the Molisch test for sugars was conducted on all samples.
-
TABLE 1 Sample Vol. Algeness ® Vol. Juvederm ® Vol. Hylenex ® Vol. β-agarase AA 200 μl-7 mg Agarose — — 200 μl-200 units AH 700 μl-24.5 mg Agarose — 700 μl-105 units — JH — 400 μl-9.6 mg HA 400 μl-60 units — In Table 1 above, the designations “AA”, “AH” and “JH” refer respectively to Algeness ®/β-agarase, Algeness ®/Hylenex ® and Juvederm ®/Hylenex ® (control) - LC/MS
- Separations were performed on an Agilent 1260 LC/MS equipped with a cooled autosampler tray (4° C.) and temperature-controlled column compartment (20° C.), which held a 2.1×100 mm i.d., 3.5 μm particle size C18 Zorbax Eclipse Plus column (Agilent, Santa Clara, Calif.). Sample injection volumes were 5 uL. A gradient elution was employed with a mobile phase composition of 0.5 mM ammonium acetate in 18Ω Millipore water (A) and acetonitrile (B), and flow rate of 0.4 mL/min. The gradient profile is shown in Table 2.
-
TABLE 2 Time (min) % A % B 0.5 100.0 0.0 5 2.0 98.0 6 2.0 98.0 7 100.0 0.0 8 100.0 0.0 - The MS was operated in positive ionization mode for agarose and negative ionization mode for hyaluronic acid, with a fragmentation voltage of 100V. Spectra were recorded in full scan mode from 300 to 2000 m/z. Neoagarotetraose, a known degradation product of agarose when exposed to β-agarase, was used to confirm enzyme activity.
- Enzyme Degradation—3.5% Agarose Gel
- Agarose gel (3.5%) was exposed to Hylenex® recombinant (hyaluronidase human injection) and β-agarase to compare the effects of enzymatic degradation. Briefly, Hylenex® (200 μl-30 units) was added to agarose (400 μl-14 mg agarose) and incubated at 37° C. for 2 hours. Additionally, β-agarase (30 μl-30 units) was added to agarose (400 μl-14 mg agarose) and incubated at 37° C. for 2 hours. Samples were centrifuged at 10000 g for 5 min at 4° C. Supernatant was filtered using 0.2 μm Nylon membrane filters and analyzed by LC/MS. Additionally, the Molisch test was conducted on all samples.
- Enzyme Degradation—Algeness® DF and Juvederm® Ultra Plus XC
- Algeness® DF (3.5% agarose) and Juvederm® Ultra Plus XC (2.4% HA) were exposed to Hylenex® recombinant (hyaluronidase human injection) to compare the effects of enzymatic degradation. In addition, Algeness® DF was also exposed to β-agarase for comparison. Briefly, enzyme was added to the dermal filler and incubated at 37° C. for 48 hours. Table 3 shows the amount of dermal filler and enzyme used for each reaction. After incubation period, samples were heated at 90° C. for 10 minutes (“kill enzyme”) and then centrifuged at 10000 g for 5 min at 4° C. Supernatant was filtered using 0.2 μm Nylon membrane filters and analyzed by LC/MS. Additionally, the Molisch test for sugars was conducted on all samples.
-
TABLE 3 Vol. Vol. Vol. Vol. β- Sample Algeness ® Juvederm ® Hylenex ® agarase AA 400 μl-14 mg — — 30 μl-30 Agarose units AH 400 μl-14 mg — 200 μl-30 — Agarose units JH — 600 μl-14.4 mg 200 μl-30 — HA units In Table 3 above, the designations “AA”, “AH” and “JH” refer respectively to Algeness ®/β-agarase, Algeness ®/Hylenex ® and Juvederm ®/Hylenex ® (control) - Results
- 0.2% Agarose Gel
- The analysis of pure agarose and β-agarase and Hylenex® were carried out to determine whether the enzymes hydrolyzed the starting material. After incubation, samples were analyzed by LC/MS to determine the presence of neoagarooligosaccharides, thus confirming if the enzyme produced the expected products. Neoagarotetraose was used as a reference standard. As expected, the analysis of the agarose/β-agarase sample produced three breakdown products, neoagarobiose, neoagarotetraose and neoagarohexaose, as shown in
FIG. 4 .FIG. 4 includes two sub-figures, the top one is an ion chromatogram for the three products represented by three peaks designated 1; 2 and 3, and the bottom one consists of three mass spectra, corresponding respectively to the three products. On the other hand, LC/MS analysis of the agarose/Hylenex® sample revealed the hexasaccharide only, as shown inFIG. 5 .FIG. 5 also includes two sub-figures, the top one is an ion chromatogram for the hexasaccharide represented by a chromatographic peak, and the bottom one is a mass spectrum corresponding to the hexasaccharide. Although results are not quantitative, neoagarotetraose produced the highest signal in the agarose/β-agarase sample. The neoagarohexaose signal from the reaction of agarose/Hylenex® was buried in the sample matrix, but observable using spectral deconvolution software, at near instrument detection limits. Based on these results, Hylenex® hydrolyzes agarose, however further studies should be conducted to determine reaction speed and amount. Note: all observations are based on one analysis and should be repeated to confirm findings. - Algeness® DF and Juvederm® Ultra Plus XC
- The Molisch test is a qualitative analysis used to indicate the presence of carbohydrates in a sample. A positive result is indicated by a purple ring between the sample and the concentrated acid. If sugars are not present in the sample, the solution will remain clear. The presence of monosaccharides results in a faster reaction, while disaccharides and polysaccharides will result in a slower reaction time.
- The Molisch test conducted on three samples: Algeness®/β-agarase, Algeness®/Hylenex® and Algeness®/water (control), contained separately in three test tubes. Immediately upon adding the Molisch reagent to each of the test tubes, the Algeness®/β-agarase sample turned dark purple, the Algeness®/Hylenex® sample remained the same, and the control (Algeness®/water) formed a light purple ring at the interface of the sample and the concentrated acid, presumably due to acid hydrolysis. After 1 hour, the Algeness®/β-agarase and the control remained the same, while the Algeness®/Hylenex® formed a light purple ring. Finally, after 2 hours, the Algeness®/Hylenex® turned a darker shade of purple. Based on these observations, the Algeness®/β-agarase presumably has smaller chain sugars present vs. Algeness®/Hylenex®. The results of the Algeness®/Hylenex® were compared to that of the Juvederm®/Hylenex®. After 19 hours, the Juvederm®/Hylenex® produced positive results (slight purple/pink). The Juvederm® control (Control J) also showed a positive result, presumably due to acid hydrolysis. When reactions are allowed to sit, the solution may turn yellow-green as observed with the Algeness® control. These samples were also analyzed by LC/MS, yielding similar results.
- Thus, the present invention has been described in detail with the understanding that the present discussion is subject to modification and alteration without departing from the teaching. Therefore, the present invention should not be limited to the precise details but should encompass the subject matter of the claims that follow and their equivalents.
Claims (12)
1-16. (canceled)
17. A drug delivery system comprising a polysaccharide and hyaluronidase held in a vessel for reconstitution.
18. The drug delivery system of claim 17 , wherein said polysaccharide has one or more sugars selected from the group consisting of D-galactose and 3, 6-anhydro-L-galactopyranose.
19. The drug delivery system of claim 18 , wherein said polysaccharide is an agarose.
20. The drug delivery system of claim 17 , further comprising one or more drugs.
21. The drug delivery system of claim 20 , wherein one or more drugs are selected from anesthetic agents, aesthetic agents, or combination thereof.
22. The drug delivery system of claim 17 , wherein the hyaluronidase render the polysaccharide in a more fluid state upon reconstitution.
23. The drug delivery system of claim 17 , wherein the polysaccharide comprises agarose and neoagarosehexaose.
24. A kit for performing dermal filling procedures comprising:
a. a polysaccharide for forming a mass in the body of an animal, wherein said polysaccharide has one or more sugars selected from the group consisting of D-galactose and 3, 6-anhydro-L-galactopyranose; and
b. an hyaluronidase for altering said polysaccharide, for administration with said polysaccharide or to said mass held in the body to effect a modification of said mass.
25. The kit of claim 24 , wherein said hyaluronidase is recombinant.
26. The kit of claim 24 , wherein said polysaccharide is an agarose.
27. The kit of claim 26 , wherein the agarose further comprises neoagarosehexaose.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US17/548,277 US20220098331A1 (en) | 2017-02-18 | 2021-12-10 | Kits and methods of using hyaluronidase to modify polysaccharide fillers and delivery systems |
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US201762460756P | 2017-02-18 | 2017-02-18 | |
PCT/US2018/000039 WO2018151827A1 (en) | 2017-02-18 | 2018-02-16 | Kits and methods of using hyaluronidase to modify polysaccharide fillers and delivery systems |
US201916486697A | 2019-08-16 | 2019-08-16 | |
US17/548,277 US20220098331A1 (en) | 2017-02-18 | 2021-12-10 | Kits and methods of using hyaluronidase to modify polysaccharide fillers and delivery systems |
Related Parent Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/US2018/000039 Division WO2018151827A1 (en) | 2017-02-18 | 2018-02-16 | Kits and methods of using hyaluronidase to modify polysaccharide fillers and delivery systems |
US16/486,697 Division US11230612B2 (en) | 2017-02-18 | 2018-02-16 | Kits and methods of using hyaluronidase to modify polysaccharide fillers and delivery systems |
Publications (1)
Publication Number | Publication Date |
---|---|
US20220098331A1 true US20220098331A1 (en) | 2022-03-31 |
Family
ID=63170431
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US16/486,697 Active 2038-05-28 US11230612B2 (en) | 2017-02-18 | 2018-02-16 | Kits and methods of using hyaluronidase to modify polysaccharide fillers and delivery systems |
US17/548,277 Abandoned US20220098331A1 (en) | 2017-02-18 | 2021-12-10 | Kits and methods of using hyaluronidase to modify polysaccharide fillers and delivery systems |
Family Applications Before (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US16/486,697 Active 2038-05-28 US11230612B2 (en) | 2017-02-18 | 2018-02-16 | Kits and methods of using hyaluronidase to modify polysaccharide fillers and delivery systems |
Country Status (4)
Country | Link |
---|---|
US (2) | US11230612B2 (en) |
EP (1) | EP3582749A4 (en) |
CN (1) | CN110719770B (en) |
WO (1) | WO2018151827A1 (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20230013351A1 (en) * | 2021-04-14 | 2023-01-19 | Veeva Systems Inc. | Computing Networks and Systems for Submitting Data |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN116056685A (en) * | 2020-02-28 | 2023-05-02 | 先进美学科技公司 | Methods, kits and compositions of novel agarose-based dermal fillers with enhanced rheological properties and safety performance |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US11235088B1 (en) * | 2018-05-04 | 2022-02-01 | Advanced Aesthetic Technologies, Inc. | Low melt temperature agarose for dermal filling and related applications and methods |
US11708462B2 (en) * | 2018-12-21 | 2023-07-25 | Advanced Aesthetic Technologies, Inc. | Crosslinked polysaccharides and related methods |
Family Cites Families (12)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE2657370C2 (en) * | 1976-12-17 | 1982-11-11 | Hans Dr.med. Dr.med.dent. 8000 München Scheicher | Means for covering and / or filling in bone defects |
CN1166407C (en) * | 2003-04-03 | 2004-09-15 | 武汉大学 | Application of agrose in preparing medicine for prostate hyperplosia |
WO2009037566A2 (en) * | 2007-06-19 | 2009-03-26 | Uvarkina Tamara P | Hyaluronidase and method of use thereof |
JP5290562B2 (en) * | 2007-10-25 | 2013-09-18 | 株式会社コーセー | Anti-wrinkle agent and skin preparation for preventing wrinkle formation |
CA2756386C (en) | 2009-03-23 | 2019-01-15 | Micell Technologies, Inc. | Drug delivery medical device |
US9408797B2 (en) * | 2011-06-03 | 2016-08-09 | Allergan, Inc. | Dermal filler compositions for fine line treatment |
US20130344048A1 (en) * | 2012-06-21 | 2013-12-26 | David H. Wasserman | Method of reducing insulin resistance by administering a Hyaluronan-degrading enzyme |
WO2014077575A1 (en) * | 2012-11-13 | 2014-05-22 | 다인바이오 주식회사 | Agar-derived neoagarooligosaccharide composite composition prepared by daga enzyme reaction and having anti-obesity and anti-diabetes effects |
ITMI20130507A1 (en) * | 2013-04-04 | 2014-10-05 | Ghimas Spa | FILLING COMPOSITION (FILLER) FOR THE TREATMENT OF LIPOATROPHIES |
US20140328826A1 (en) * | 2013-05-01 | 2014-11-06 | COLE Research & Design, Inc. | Product and method for treating keloid scars, hypertrophic scars and burn scars with contracture |
WO2016100476A1 (en) * | 2014-12-16 | 2016-06-23 | KeraNetics, LLC | Dermal fillers comprising keratin biomaterials |
EP3344064A4 (en) * | 2015-03-24 | 2019-08-07 | Advanced Aesthetic Technologies, Inc. | Kits and methods of using enzymes to modify polysaccharide fillers and delivery systems |
-
2018
- 2018-02-16 EP EP18754752.6A patent/EP3582749A4/en active Pending
- 2018-02-16 US US16/486,697 patent/US11230612B2/en active Active
- 2018-02-16 CN CN201880019605.1A patent/CN110719770B/en active Active
- 2018-02-16 WO PCT/US2018/000039 patent/WO2018151827A1/en unknown
-
2021
- 2021-12-10 US US17/548,277 patent/US20220098331A1/en not_active Abandoned
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US11235088B1 (en) * | 2018-05-04 | 2022-02-01 | Advanced Aesthetic Technologies, Inc. | Low melt temperature agarose for dermal filling and related applications and methods |
US11708462B2 (en) * | 2018-12-21 | 2023-07-25 | Advanced Aesthetic Technologies, Inc. | Crosslinked polysaccharides and related methods |
Non-Patent Citations (1)
Title |
---|
Temuujin et al. Overexpression and biochemical characterization of DagA from Streptomyces coelicolor A3(2): an endo-type β-agarase producing neoagarotetraose and neoagarohexaose. Appl Microbiol Biotechnol (2011) 92:749–759. (Year: 2011) * |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20230013351A1 (en) * | 2021-04-14 | 2023-01-19 | Veeva Systems Inc. | Computing Networks and Systems for Submitting Data |
US11811522B2 (en) * | 2021-04-14 | 2023-11-07 | Veeva Systems Inc. | Computing networks and systems for submitting data |
Also Published As
Publication number | Publication date |
---|---|
WO2018151827A8 (en) | 2018-10-25 |
WO2018151827A9 (en) | 2018-11-29 |
WO2018151827A1 (en) | 2018-08-23 |
US20200010577A1 (en) | 2020-01-09 |
US11230612B2 (en) | 2022-01-25 |
CN110719770A (en) | 2020-01-21 |
EP3582749A1 (en) | 2019-12-25 |
EP3582749A4 (en) | 2020-12-30 |
CN110719770B (en) | 2024-04-02 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US20220098331A1 (en) | Kits and methods of using hyaluronidase to modify polysaccharide fillers and delivery systems | |
CN102574902B (en) | SAP derivative and preparation thereof and purposes | |
CN102238953B (en) | Be used for the treatment of the injectable poly-deoxyribonucleotide compositions of osteoarthritis | |
EP2540284A2 (en) | Compositions and methods for stabilized polysaccaride formulations | |
EP3053604B1 (en) | Compositions and methods for treating joints | |
JP2007238627A (en) | Therapeutic agent comprising botulinum neurotoxin | |
MX2014004792A (en) | Method of treating or reducing efp. | |
KR20140057824A (en) | Stabilizer for hyaluronidase and liquid formulation comprising hyaluronidase | |
JP2016523916A (en) | Stable liquid formulation | |
AU2004224510B2 (en) | Remedy for nerve damage | |
US10668091B2 (en) | Antibacterial agent against multidrug-resistant gram-positive bacteria and external agent | |
JP2012062273A (en) | Glycogen-containing agent for promoting production of hyaluronic acid and elastin | |
Matarasso et al. | Hypersensitivity reaction to nonanimal stabilized hyaluronic acid | |
JP2013170149A (en) | Extracellular substrate formation-promoting agent containing hyaluronic acid oligosaccharide as active ingredient | |
AU2002327105B2 (en) | Separate type medical material | |
CN109806228A (en) | A kind of solution containing meloxicam liposome | |
US20210228468A1 (en) | Kits and methods of using ascorbates to modify polysaccharide fillers and delivery systems | |
EP2578228B1 (en) | Non-diffusive botulinum toxin causing local muscle paralysis, and purification method thereof | |
EP3344064A1 (en) | Kits and methods of using enzymes to modify polysaccharide fillers and delivery systems | |
JP4451070B2 (en) | CD44 cleavage inducer | |
RU2741948C1 (en) | Method for preventing hypercoagulation in animals under stress | |
Kang et al. | ENHANZE® Drug Delivery Technology: Advancing Subcutaneous Drug Delivery Using Recombinant Human Hyaluronidase PH20 | |
CA3132923A1 (en) | Stable pharmaceutical formulations of peptide and protein drugs | |
CN115671267A (en) | Subcutaneous antibiotic pharmaceutical composition | |
CN103221066A (en) | Sustained-release pharmaceutical composition |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
STPP | Information on status: patent application and granting procedure in general |
Free format text: DOCKETED NEW CASE - READY FOR EXAMINATION |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: NON FINAL ACTION MAILED |
|
STCB | Information on status: application discontinuation |
Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION |