WO2013071216A1 - Matière de remplissage injectable - Google Patents

Matière de remplissage injectable Download PDF

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Publication number
WO2013071216A1
WO2013071216A1 PCT/US2012/064586 US2012064586W WO2013071216A1 WO 2013071216 A1 WO2013071216 A1 WO 2013071216A1 US 2012064586 W US2012064586 W US 2012064586W WO 2013071216 A1 WO2013071216 A1 WO 2013071216A1
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WIPO (PCT)
Prior art keywords
cross
polymer
linked
phase
gel
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PCT/US2012/064586
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English (en)
Inventor
Phi Nguyen
Loc Phan
Bao Tran
Thuan Nguyen
Original Assignee
Miba Medical Inc.
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Publication date
Application filed by Miba Medical Inc. filed Critical Miba Medical Inc.
Priority to CN201280050924.1A priority Critical patent/CN103917256A/zh
Priority to KR1020157025171A priority patent/KR20150111372A/ko
Priority to KR1020147007016A priority patent/KR20140059238A/ko
Priority to JP2014526281A priority patent/JP2014521492A/ja
Priority to EP12847290.9A priority patent/EP2776077A4/fr
Priority to US13/872,071 priority patent/US20140256695A1/en
Publication of WO2013071216A1 publication Critical patent/WO2013071216A1/fr

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    • 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
    • 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
    • A61L27/26—Mixtures of macromolecular compounds
    • 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
    • A61L27/52—Hydrogels or hydrocolloids
    • 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
    • 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
    • A61L27/58—Materials at least partially resorbable by the body
    • 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
    • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
    • A61L31/04—Macromolecular materials
    • A61L31/041—Mixtures of macromolecular compounds
    • 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
    • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
    • A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • 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
    • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
    • A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L31/145—Hydrogels or hydrocolloids
    • 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
    • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
    • A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L31/148—Materials at least partially resorbable by the body
    • 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
    • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
    • A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L31/16—Biologically active materials, e.g. therapeutic substances
    • 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
    • C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
    • 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
    • C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
    • C08B37/0063—Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan
    • C08B37/0072—Hyaluronic acid, i.e. HA or hyaluronan; Derivatives thereof, e.g. crosslinked hyaluronic acid (hylan) or hyaluronates
    • 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
    • A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
    • A61L2300/402—Anaestetics, analgesics, e.g. lidocaine
    • 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
    • A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
    • A61L2300/41—Anti-inflammatory agents, e.g. NSAIDs
    • 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
    • A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
    • A61L2300/43—Hormones, e.g. dexamethasone
    • 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
    • A61L2300/60—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a special physical form
    • A61L2300/602—Type of release, e.g. controlled, sustained, slow
    • A61L2300/604—Biodegradation
    • 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
    • A61L2430/02—Materials or treatment for tissue regeneration for reconstruction of bones; weight-bearing implants
    • 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
    • A61L2430/04—Materials or treatment for tissue regeneration for mammary reconstruction
    • 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

Definitions

  • the present invention relates to biocompatible viscoelastic polymeric gel slurries, methods for their preparation and formulations containing them.
  • systems and methods are disclosed for cosmetic augmentation of soft tissue using cross- linked HA that had been optimized for
  • IPN interpenetrating network
  • the IPN configuration gives this cross linked HA those utilities unique for this cosmetic augmentation application.
  • the IPN core (imagine a tapioca ball) is more resistance to biodegradation in a human body than the single cross-linked material normalized for the same cross linking level.
  • varying physical properties that continuously changes radiating out from the core makes the polymer tough and at the same time compliant with the local tissue for better tissue/device biocompatibility and feels more natural to the touch.
  • the above HA cross linking method optimized for cosmetic augmentation in certain cases may need to control delivered pharmaceutical substances to modulate local tissue response to the polymer.
  • the pharmaceutical component makes up the multi-phase mixture with the other phase being the cross linked HA polymer
  • Implementations of the above aspects may include one or more of the following.
  • the system is biocompatible and performs controlled drug releases at strategic timing to coincide with key physiological events. For example, a fast drug release profile and no delay would be well suited for the controlled release of an anesthetic such as lidocane to relieve acute pain experienced by the patient associated with the surgical procedure.
  • the system is also capable of a medium release profile and a medium delay of a corticosteroid or steroid such as dexamethasone or triamcinolone to co-inside with a physiological inflammatory foreign body reaction.
  • the system can also be customized to have a medium to slow release profile and a longer delay before starting the release of an antiproliferative drug such as paclitaxel, serolimas or 5-flourouracil to stop uncontrolled healing and excessive remodeling causing unsightly scar formation or capsular formation.
  • an antiproliferative drug such as paclitaxel, serolimas or 5-flourouracil to stop uncontrolled healing and excessive remodeling causing unsightly scar formation or capsular formation.
  • Another aspect of the present invention includes methods for optimizing biodegradation profiles and control migration of the implant material through the manipulation of various types molecular weight.
  • the system optimizes biodegradation profiles and controls migration of the implant material.
  • the system can be formulated around various types of molecular weights such as M n , M w and M z, and their polydispersity index (PDI) to optimize the biodegradation profiles to be from hypervolumic to isovolumic to hypovolumic.
  • PDI polydispersity index
  • HA in the body is biodegraded by twomajor mechanisms: oxidative and hydrolytic.
  • the mechanism is enzymatic hydrolysis by three enzymes hyaluronidase (hyase), b-d-glucuronidase, and ⁇ - ⁇ - acetyl-hexosaminidase, and outside the cell the mechanism is oxidation by oxygen derived free radical, or sometimes, they are called reactive oxygen species (ROS).
  • ROS reactive oxygen species
  • ROS are produced as a normal product of cellular metabolism.
  • one major contributor to oxidative damage is hydrogen peroxide (H202), which is converted from superoxide that leaks from the mitochondria.
  • Catalase and superoxide dismutase ameliorate the damaging effects of hydrogen peroxide and superoxide by converting these compounds into oxygen and water, benign molecules.
  • this conversion is not 100% efficient, and residual peroxides persist in the cell.
  • ROS are produced as a product of normal cellular functioning, excessive amounts can cause deleterious effects. Memory capabilities decline with age, evident in human degenerative diseases such as Alzheimer's disease, which is accompanied by an accumulation of oxidative damage.
  • Older gerbils were found to have higher levels of oxidized protein in comparison to younger gerbils. Treatment of old and young mice with a spin trapping compound caused a decrease in the level of oxidized proteins in older gerbils but did not have an effect on younger gerbils. In addition, older gerbils performed cognitive tasks better during treatment but ceased functional capacity when treatment was discontinued, causing oxidized protein levels to increase.
  • the degradation reaction by oxygen derived free radical of HA was the results of studies using the HA present in synovial fluids. It showed that the HA was readily degraded by super oxide free radicals. This reaction is most favorable in the case of secondary free radicals.
  • Neutrophils polymorphonuclear leukocytes
  • These WBC's are by far the exclusive destroyers of HA by oxygen-derived free radical mechanism.
  • an aspect of this invention is to quench the effect of the free radical before it degrades the HA using free radical scavengers such as antioxidant vitamins.
  • Antioxidants are intimately involved in the prevention of cellular damage—the common pathway for cancer, aging, and a variety of diseases. Antioxidants are molecules which can safely interact with free radicals and terminate the chain reaction before vital molecules are damaged. Although there are several enzyme systems within the body that scavenge free radicals, the principle micronutrient (vitamin) antioxidants are vitamin E, beta-carotene, and in the case of HA, vitamin C is the exception. Additionally, selenium, a trace metal that is required for proper function of one of the body's antioxidant enzyme systems, is sometimes included in this category. The body cannot manufacture these micronutrients so they must be supplied in the diet.
  • Vitamin E d-alpha tocopherol.
  • a fat soluble vitamin present in nuts, seeds, vegetable and fish oils, whole grains (esp. wheat germ), fortified cereals, and apricots.
  • Current recommended daily allowance (RDA) is 15 IU per day for men and 12 IU per day for women.
  • Vitamin C The exception in the case of HA as it is detrimental to the longevity of HA.
  • vitamin C is ascorbic acid, and it is a water soluble vitamin present in citrus fruits and juices, green peppers, cabbage, spinach, broccoli, kale, cantaloupe, kiwi, and strawberries.
  • the RDA is 60 mg per day. Intake above 2000 mg may be associated with adverse side effects in some individuals.
  • Vitamin A is a precursor to vitamin A (retinol) and is present in liver, egg yolk, milk, butter, spinach, carrots, squash, broccoli, yams, tomato, cantaloupe, peaches, and grains. Because beta-carotene is converted to vitamin A by the body there is no set requirement. Instead the RDA is expressed as retinol equivalents (RE), to clarify the relationship. (NOTE: Vitamin A has no antioxidant properties and can be quite toxic when taken in excess.)
  • Glutathione is a tripeptide with a gamma peptide linkage between the amine group of cysteine (which is attached by normal peptide linkage to a glycine) and the carboxyl group of the glutamate side-chain. It is an antioxidant, preventing damage to important cellular components caused by reactive oxygen species such as free radicals and peroxides. Thiol groups are reducing agents, existing at a concentration of approximately 5mM in animal cells. Glutathione reduces disulfide bonds formed within cytoplasmic proteins to cysteines by serving as an electron donor. In the process, glutathione is converted to its oxidized form glutathione disulfide (GSSG), also called L(-)-Glutathione.
  • GSSG glutathione disulfide
  • glutathione can be reduced back by glutathione reductase, using NADPH as an electron donor.
  • the ratio of reduced glutathione to oxidized glutathione within cells is often used as a measure of cellular toxicity.
  • Uric Acid It is the most important plasma antioxidant in humans, and a heterocyclic compound of carbon, nitrogen, oxygen, and hydrogen with the formula C5H4N403. It forms ions and salts known as urates and acid urates such as ammonium acid urate. Uric acid is a product of the metabolic breakdown of purine nucleotides. High blood concentrations of uric acid can lead to a type of arthritis known as gout. The chemical is associated with other medical conditions including diabetes and the formation of ammonium acid urate kidney stones.
  • antioxidant enzymes to protect the longevity of HA. These enzymes can reduce the radicals and defend against ROS. They are: alpha- 1- microglobulin, superoxide dismutases, catalases, lactoperoxidases, glutathione peroxidases and peroxiredoxins.
  • an aspect of this invention uses hyaluronidase inhibitor (anti-HA) to prevent the depolymerization of HA, specifically by hyaluronidase, and to maintain the longevity of HA. Maintenance of HA longevity is important because it is directly related to the appearance of those unwanted wrinkles and the signs of aging.
  • anti-HA hyaluronidase inhibitor
  • HA is an important molecule to everything that lives on this earth. In that, it is a multifunctional high molecular weight polysaccharide found throughout the animal kingdom, especially in the extracellular matrix (ECM) of soft connective tissues. HA is thought to participate in many biological processes, and its level is markedly elevated during embryogenesis, cell migration, wound healing, malignant transformation, and tissue turnover. The enzymes that degrade HA, hyaluronidases (HAases) are expressed both in prokaryotes and eukaryotes. These enzymes are known to be involved in physiological and pathological processes ranging from fertilization to aging.
  • ECM extracellular matrix
  • Hyaluronidase-mediated degradation of HA increases the permeability of connective tissues and decreases the viscosity of body fluids and is also involved in bacterial pathogenesis, the spread of toxins and venoms, acrosomal reaction/ovum fertilization, and cancer progression. Furthermore, these enzymes may promote direct contact between pathogens and the host cell surfaces. Depolymerization of HA also adversely affects the role of ECM and impairs its activity as a reservoir of growth factors, cytokines and various enzymes involved in signal transduction. Inhibition of HA degradation therefore may be crucial in reducing disease progression and spread of venom/toxins and bacterial pathogens.
  • Hyaluronidase inhibitors are potent, ubiquitous regulating agents that are involved in maintaining the balance between the anabolism and catabolism of HA. Hyaluronidase inhibitors could also serve as contraceptives and anti-tumor agents and possibly have antibacterial and anti-venom/toxin activities. Additionally, these molecules can be used as pharmacological tools to study the physiological and pathophysiological role of HA and hyaluronidases.
  • Alkaloids Aristolochic acid, ajmaline. reseipine
  • Synthetic PS53 (Mydroquinone-sulfonic acid-formaldehyde polymer) ⁇ phosphory!ated hespei din, polymer of compounds poly (styrene-4-sulfonate), sodium cellulose sulfate, 1 -tetradecane sulfonic acid, L-arginin derivatives, traxanox, nor!ignane, urolithin B, aescin, dipiienylacrylie acids, diphenyl propionic acids, indole derivatives, ehalcone derivatives.
  • Glycosaminogiycans Heparin, heparan sulfate, beatan sulfate, chondroirin sulfate (A, C, D), O-siiifated ⁇ , linamarin, and glycosides. amygdalin.
  • FIG. 1 shows an exemplary system to produce multiply cross-linked HA.
  • FIG. 2 shows another exemplary system to produce multiply cross-linked HA.
  • FIG. 3 shows an exemplary diagram of the resulting multiply cross-linked HA.
  • the hyaluronan of a recombinant Bacillus cell is expressed directly to the culture medium, a simple process may be used to isolate the hyaluronan from the culture medium.
  • the Bacillus cells and cellular debris are physically removed from the culture medium.
  • the culture medium may be diluted first, if desired, to reduce the viscosity of the medium.
  • Many methods are known to those skilled in the art for removing cells from culture medium, such as centrifugation or microfiltration. If desired, the remaining supernatant may then be filtered, such as by ultrafiltration, to concentrate and remove small molecule contaminants from the hyaluronan.
  • a simple precipitation of the hyaluronan from the medium is performed by known mechanisms.
  • Salt, alcohol, or combinations of salt and alcohol may be used to precipitate the hyaluronan from the filtrate.
  • the hyaluronan can be easily isolated from the solution by physical means.
  • the hyaluronan may be dried or concentrated from the filtrate solution by using evaporative techniques known to the art, such as lyophilization or spray drying.
  • the content of hyaluronic acid may be determined according to the modified carbazole method (Bitter and Muir, 1962, Anal Biochem. 4: 330-334). Moreover, the number average molecular weight of the hyaluronic acid may be determined using standard methods in the art, such as those described by Ueno et al, 1988, Chem. Pharm. Bull . 36, 4971-4975; Wyatt, 1993, Anal. Chim. Acta 272: 1-40; and Wyatt Technologies, 1999, "Light Scattering University DAWN Course Manual” and "DAWN EOS Manual” Wyatt Technology Corporation, Santa Barbara, Calif.
  • the hyaluronic acid, or salt thereof, of the one embodiment has a molecular weight of about 10,000 to about 10,000,000 Da. In a more preferred embodiment it has a molecular weight of about 25,000 to about 5,000,000 Da. In a most preferred embodiment, the hyaluronic acid has a molecular weight of about 50,000 to about 3,000,000 Da. In another embodiment, the hyaluronic acid or salt thereof has a molecular weight in the range of between 300,000 and 3,000,000; preferably in the range of between 400,000 and 2,500,000; more preferably in the range of between 500,000 and 2,000,000; and most preferably in the range of between 600,000 and 1,800,000.
  • the hyaluronic acid or salt thereof has a low number average molecular weight in the range of between 10,000 and 800,000 Da; preferably in the range of between 20,000 and 600,000 Da; more preferably in the range of between 30,000 and 500,000 Da; even more preferably in the range of between 40,000 and 400,000 Da; and most preferably in the range of between 50,000 and 300,000 Da.
  • This example illustrates the preparation of DVS-crosslinked microparticles.
  • Sodium hyaluronate (HA, 580 kDa, 1.90 g) was dissolved in aqueous NaOH (0.2 M, 37.5 ml) by vigorous stirring at room temperature for 3 hours until a homogenous solution was obtained.
  • Sodium chloride (0.29 g) was added and mixed shortly.
  • Mineral oil (10.0 g) and ABIL® EM 90 surfactant (Cetyl PEG/PPG-10/1 Dimethicone, 1.0 g) were mixed by stirring.
  • Divinylsulfone (DVS, 320 microliter) was added to the aqueous alkaline HA-solution and mixed for 1 min. to obtain a homogeneous distribution in the aq. phase.
  • the water phase was then added within 2 minutes to the oil phase with mechanical stirring at low speed.
  • An emulsion was formed immediately and stirring was continued for 30 minutes at room temperature.
  • the emulsion was left over night at room temperature.
  • the emulsion was neutralized to pH 7.0 by addition of aq. HCI (4 M, approx. 2.0 ml) and stirred for approx. 40 min.
  • This example illustrates the preparation of DVS-crosslinked microparticles with neutralization using a pH indicator.
  • Sodium hyaluronate (HA, 580 kDa, 1.88 g) was dissolved in aqueous NaOH (0.2 M, 37.5 ml) by vigorous stirring at room temperature for 2 hours until a homogenous solution was obtained.
  • Bromothymol blue pH indicator (equivalent range pH 6.6-6.8) was added (15 drops, blue color in solution).
  • Sodium chloride (0.25 g) was added and mixed shortly.
  • Divinylsulfone (DVS, 320 microliter) was added to the aqueous alkaline HA-solution and mixed very vigorously for 30 to 60 seconds to obtain a homogeneous distribution in the aq. phase.
  • the water phase was then added within 30 sec. to the oil phase with mechanical stirring at 400 RPM.
  • An emulsion was formed immediately and stirring was continued for 30min. at room temperature.
  • Neutralization was performed by addition of aq. HCI (4 M, 1.6 ml) and the emulsion was left at room temperature with magnetic stirring for 4 hours.
  • the pH indicator present in the gel particles changed color to green. pH in the emulsion was measured by pH stick to 3-4. The emulsion was left in fridge overnight. The pH indicator present in the gel particles had changed to yellow.
  • This example illustrates the breakage of the W/O emulsion followed by phase separation and dialysis.
  • the crosslinked HA microparticles were separated from the W/O emulsion by organic solvent extraction.
  • the W/O emulsion (5 g) and a mixture of n-butanol/chloroform (1/1 v%, 4.5 ml) was mixed vigorously by whirl mixing in a test tube at room temperature. Extra mQ-water (20 ml) was added to obtain phase separation.
  • the test tube was centrifuged and three phases were obtained with the bottom phase being the organic phase, middle phase of gel particles and upper phase of clear aqueous solution.
  • This example illustrates the preparation of DVS-crosslinked HA microparticles.
  • Sodium hyaluronate (HA, 580 kDa, 1.89 g) was dissolved in aqueous NaOH (0.2 M, 37.5 ml).
  • Sodium chloride (0.25 g) was added and the solution was stirred by magnetic stirring for 1 hour at room temperature until a homogeneous solution was obtained.
  • TEGOSOFT® M (10.0 g) oil and ABIL® EM 90 surfactant (Cetyl PEG/PPG- 10/1 Dimethicone, 1.0 g) were mixed by stirring.
  • Divinylsulfone (DVS, 320 microliter) was added to the aqueous alkaline HA-solution and mixed for 1 min. to obtain a homogenoues distribution in the aq. phase.
  • the water phase was then added within 2 min. to the oil phase with mechanical stirring (300 RPM). An emulsion was formed immediately and stirring was continued for 30 min. at room temperature.
  • the emulsion was neutralized by addition of stociometric amounts of HCI (4 M, 1.8 ml) and stirred for approx. 40 min.
  • the emulsion was broken by addition of a n-butanol/chloroform mixture (1 : 1 v%, 90 ml) and extra MilliQ®-water (100 ml) followed by magnetic stirring.
  • the upper phase was separated in a volume of approx. 175 ml.
  • the organic phase was mixed with mQ-water (30 ml) for a final washing.
  • the combined water/gel phase (205 ml) were transferred to a dialysis tube (MWCO 12-14,000, Diameter 29 mm, Vol/Length 6.4 ml/cm) and dialysed against MilliQ®-water overnight at room temperature.
  • the conductivity were decreased to 0.67 micro-Sievert/cm after subsequent change of water (3 times) and dialysis overnight (2 nights).
  • the microparticles were assessed by microscopy (DIC 200x), see FIG. 1; the cross-section of one microparticle is indicated and labelled "21,587.92 nm".
  • This example illustrates the breakage of the W/O emulsion and isolation of the gel microparticles.
  • the gel microparticles were separated from the W/O-emulsion by organic extractions.
  • organic solvents which were used for this extraction were mixtures of butanol/chloroform in volume ratios (v%) of 75:20 to 20.80, respectively.
  • the weight ratio (w%) of W/O emulsion to organic solvent was approximately 1 : 1.
  • Separation in small scale The W/O emulsion (5 g) was weighed in centrifuge tubes (50 ml). A mixture of butanol/ chloroform was prepared (1 : 1 v%) and from this mixture 4.5 ml was added (corresponds to 5 g) to the test tube.
  • test tube was carefully mixed to secure that all emulsion was dissolved.
  • the test tube was mixed by Whirl mixing and left at room temperature for phase separation. Phase separation with water phase on top and organic phase at bottom with a white emulsion phase in between was often observed. Addition of more water and organic phases improved separation.
  • the water phase was separated by decanting and further purified or characterized.
  • This example illustrates a composition in which the HA microparticles were formed.
  • a hot/cold procedure can be used with incorporation of a cold water phase B into a hot oil phase, which will shorten the time of manufacture.
  • a non-limiting example of formulation could be as follows:
  • Phase B Solubilize hyaluronic acid (Hyacare®) in aq. NaOH by stirring; then add NaCl and stir.
  • HA hyaluronate
  • aqueous NaOH 0.2 M, 37.5 mL
  • Sodium chloride 0.25 g
  • surfactant ABIL® EM 90
  • VS Divinylsulfone
  • the water phase was then added within 2 min to the oil phase with mechanical stirring (300 RPM). An emulsion was formed immediately and stirring was continued for 30 min at room temperature.
  • the emulsion was neutralized by addition of stociometric amounts of HCI (4 M, 1.8 mL) and stirred for approx. 40 min.
  • the emulsion was transferred to a separation funnel, and broken by addition of a n-butanol/chloroform mixture (1 : 1 v%, 90 mL) and extra millliQTM-water (100 mL) followed by vigorous shaking.
  • the upper phase was separated in a volume of approx. 175 mL.
  • the organic phase was washed with millliQTM-water (100 mL).
  • the combined water/gel phase was transferred to a dialysis tube (MWCO 12-14,000, Diameter 29 mm, Vol/Length 6.4 mL/cm) and dialysed against millliQTM-water overnight at room temperature.
  • the conductivity was decreased to 10 micro-Sievert/cm after subsequent change of water (3 times) and dialysis overnight (2 nights).
  • This example illustrates the final isolation and purification of the microparticles.
  • This example illustrates performance of rheological studies on particles.
  • a particle sample is analyzed on an Anton Paar rheometer (Anton Paar GmbH, Graz, Austria, Physica MCR 301, Software: Rheoplus), by use of a 50 mm 2° cone/plate geometry.
  • First the linear range of the visco-elastic properties G' (Storage modulus) and G" (Loss modulus) of the material is determined by an amplitude sweep with variable strain, ⁇ . Secondary a Frequency sweep is made, and based on values of the visco-elastic values, G' and G", tan ⁇ can be calculated as a value for week/strong gel behaviors.
  • This example illustrates performance of an investigation of force applied to inject at a certain speed, as a function of the homogeneity of the sample.
  • a particle sample is transferred to a syringe applied with a needle, either 27Gx1 ⁇ 2 ", 30Gx1 ⁇ 2 ", and is set in a sample rig, in a texture analyzer (Stable Micro Systems, Surrey, UK, TA.XT Plus, Software: Texture Component 32).
  • the test is performed with an injection speed at 12 . 5 mm/min., over a given distance.
  • This example illustrates the preparation of DVS-cross-linked HA hydrogels with concomitant swelling and pH adjustment.
  • the pH of the gels was stabilized during the swelling step. After swelling, any excess buffer was removed by filtration and the hydrogels were briefly homogenized with an IKA® ULTRA-TURRAX® T25 homogenizer (Ika Labortechnik, DE). The volume and pH of the gels were measured (see Table 2).
  • This example illustrates the preparation of highly homogenous DVS-cross-linked HA hydrogels.
  • Sodium hyaluronate (770 kDa, 2 g) was dissolved into 0.2M NaOH with stirring for approx. 1 hour at room temperature to give a 8% (w/v) solution. DVS was then added so that the HA/DVS weight ratio was 7: 1. After stirring at room temperature for 5 min, one of the samples was heat treated at 50° C. for 2 h without stirring, and then allowed to stand at room temperature overnight. The resulting cross-linked gel was swollen into 200 ml phosphate buffer (pH 5.5) 37° C. for 42 or 55 h, and finally washed twice with 100 ml water, which was discarded. Volume and pH were measured, as well as the pressure force necessary to push the gels through a 27G*1 ⁇ 2 injection needle (see Table 3).
  • the cross-linked HA hydrogel prepared according to this example exhibited a higher swelling ratio and an increased softness compared to a control hydrogel which was not heat treated (Table 3).
  • the pressure force applied during injection through a 27G*1 ⁇ 2 needle was more stable than that of the latter sample, indicating that the cross-linked HA hydrogel is more homogenous.
  • This example illustrates the in vitro biostability of DVS-cross-linked HA hydrogels using enzymatic degradation.
  • a bovine testes hyaluronidase (HAase) solution (100 U/mL) was prepared in 30 mM citric acid, 150 mM Na 2 HPO 4 , and 150 mM NaCl (pH 6.3).
  • DVS-HA cross-linked hydrogel samples (ca. 1 mL) were placed into safe-lock glass vials, freeze-dried, and weighed (W 0 ; Formula 1).
  • the enzyme solution (4 mL, 400 U) was then added to each sample and the vials were incubated at 37° C. under gentle shaking (100-200 rpm). At predetermined time intervals, the supernatant was removed and the samples were washed thoroughly with distilled water to remove residual salts, they were then freeze-dried, and finally weighed (W t ; Formula 1).
  • the biodegradation is expressed as the ratio of weight loss to the initial weight of the sample (Formula 1). Weight loss was calculated from the decrease of weight of each sample before and after the enzymatic degradation test. Each biodegradation experiment was repeated three times. DVS-HA hydrogels prepared as described in example 2 ('Heated') were compared to DVS-HA hydrogels which had not been heat treated ('Not heated'). For both types of gel, degradation was fast during the first four hours, and then proceeded slower until completion at 24 h. Importantly there was a significant variation of the weight loss values for the samples which had not been heated as compared to the hydrogel prepared with a heating step as described in example 2. This clearly illustrates that a highly homogenous DVS-cross-linked HA hydrogel is obtained by using the process described in example 2.
  • DVS-crosslinked HA hydrogels were formulated into creams and serums, that when applied to the skin increase the skin moisturization and elasticity, and provide immediate anti-aging effect, as well as film-forming effect
  • phase E A typical formulation of a water-in-oil (w/o) emulsion containing 2% DVS-cross-linked HA.
  • Each phase (A to E) was prepared separately by mixing the defined ingredients (see Table 4).
  • Phase B was then added to phase A under stirring with a mechanical propel stirring device and at a temperature less than 40° C.
  • Phase C was then added followed by phase D and finally phase E under stirring.
  • Formulations were also made, wherein the HA hydrogel concentration was 4%, 6% and 8%, respectively, in Phase D, to give a range of w/o formulations.
  • phase E Another typical formulation of a w/o-emulsion containing 2% DVS-crosslinked HA is shown in table 5.
  • Each phase (A to F) in table 5 was prepared separately by mixing the defined ingredients (see Table 5).
  • Phase B was mixed with phase A and the resulting oil phase was heated at 75° C.
  • Phase C was also heated to 75° C.
  • the oil phase was added to phase C at 75° C. under stirring with a mechanical propel stirring device.
  • the emulsion was then cooled down to less than 40° C, after which phase D was added, followed by phase E and finally phase F under stirring.
  • Formulations were also made, wherein the HA hydrogel concentration was 4%, 6%> and 8%, respectively, in Phase E, to give a range of w/o formulations.
  • a typical formulation of a silicone serum containing 2% DVS-cross-linked HA was prepared as shown in table 6. All ingredients were mixed at the same time under very high stirring and at less than 40° C. (see table 6). Formulations were also prepared, wherein the HA hydrogel concentration was 4%, 6%> and 8%, respectively, to give a range of serums. TABLE 6
  • DVS cross-linked HA hydrogels with neutral pH are obtained after swelling in phosphate buffer (pH 7.0) for 8 to 14 hours, depending on the degree of cross-linking.
  • a set of DVS cross-linked HA hydrogels was prepared as described in the above, using from 4 to 8% HA solution, and using various amounts of DVS cross-linker, as indicated in Table 7.
  • the decrease in pH is shown for the HA 6% solution and two different ratios of HA/DVS in FIG. 2, where the HA/DVS ratio of 10:1 is labelled with triangles, and 15: 1 is labelled with squares. In these two cases, pH was neutralized within 8 hours. In contrast, neutral pH was reached after 14 hour-swelling for hydrogels with either a higher HA concentration (e.g. 8%) or a higher degree of cross- linking (e.g. HA/DVS ratio of 2.5).
  • Example 17 Visco Elastic Properties of Hydrogels Based on DVS-Crosslinked HA
  • the rheological measurements were performed on a Physica MCR 301 rheometer (Anton Paar, Ostfildern, Germany) using a plate-plate geometry and at a controlled temperature of 25° C.
  • the visco-elastic behavior of the samples was investigated by dynamic amplitude shear oscillatory tests, in which the material was subjected to a sinusoidal shear strain.
  • strain/amplitude sweep experiments were performed to evaluate the region of deformation in which the linear viscoelasticity is valid.
  • the strain typically ranged from 0.01 to 200% and the frequency was set to 1 Hz.
  • the shear storage modulus (or elastic modulus G') and the shear loss modulus (or viscous modulus, G") values were recorded from frequency sweep experiments at a constant shear strain (10%) and at a frequency between 0.1 and 10 Hz.
  • the geometry, the NF and the gap were PP 25, 2 and 1 mm, respectively.
  • G' gives information about the elasticity or the energy stored in the material during deformation
  • G" describes the viscous character or the energy dissipated as heat.
  • the elastic modulus gives information about the capability of the sample to sustain load and return in the initial configuration after an imposed stress or deformation. In all experiments, each sample was measured at least three times.
  • a DVS-cross-linked HA hydrogel was prepared using 1.5 g of sodium HA in 0.2 M NaOH to give a 6% (w/v) solution.
  • the HA/DVS weight ratio was 10: 1.
  • the hydrogel was prepared in three replicates according to the procedure described in example 2 until the swelling step, after which it was treated as follows: After incubation in an oven at 50° C. for two hours, the hydrogel was immersed into Na2HP04/NaH2P04 buffer (1 L, 50 mM, pH 7.0) containing the preservative (2-phenoxyethanol/3 [(2-ethylhexyl)oxy] 1 ,2-propanediol).
  • the concentration of preservative was 10 mL/mL to target a final concentration of 1% (v/v) in the swollen hydrogel. It was anticipated that the preservative would diffuse into the hydrogel during the incubation, and that at the same time, microbial contamination in the buffer would be prevented.
  • the vessel was covered with parafilm and placed in an oven at 37° C. After 1 h, the swelling bath was removed and the hydrogel was swollen in a fresh phosphate buffer containing 10 mL/mL preservative for 6-7 h. This step was repeated until the swelling time was 12 h, whereafter the pH was measured. Swelling was continued for another 2.5 h to reach neutral pH.
  • the amount of preservative incorporated into the hydrogel was determined by UV- spectrophotometry (Thermo Electron, Nicolet, Evolution 900, equipment nr. 246-90). A 1% (v/v) solution of the preservative in phosphate buffer was first analyzed to select the wavelength. Approximately 5 mL of hydrogel were collected using a pipette. Typically, samples were collected in the center of the swollen round hydrogel, and in the north, east, south, and west "sides" of the round gel.
  • the samples were then transferred into a cuvette and the absorbance was read at 292 nm. Each sample was read three times and the absorbance was zeroed against a blank DVS-cross-linked HA hydrogel, containing no preservative.
  • the time of degradation may be adjusted based on the polymer mixture in Table 1 below.
  • Examples 1 and 2 below are examples of matrix incorporation of drug or drugs into a biodegradable polymer to control the releases the drugs.
  • biodegradable polymer may be used to control the degradation timing and / or to control the degradation by-products.
  • Some biodegradable polymers are:
  • PGA Poly(glycolic acid)
  • PLA poly(lactic acid)
  • the particle sizes of the micro capsules are directly controlled by the interfacial chemistry of the organic phase and the aqueous phase.
  • a surfactant is often used to mediate interfacial surface chemistry between an oily substance and the aqueous environment.
  • a surfactant is a detergent that is in an aqueous solution.
  • Surfactants are large molecules that have both polar and non-polar ends. The polar end of the molecule will attach itself to water, also a polar molecule. The non-polar end of the molecule will attract NAPL (non-aqueous phase liquid) compounds.
  • surfactants that are used for solubilization are:
  • Sioponic 25-9 which is a linear alcohol ethoxylate, and has a solubilization value of 2.75g/g
  • Tergitol which is an ethylene oxide / propylene oxide with a solubilization value of 1.21g/g
  • Tergitol XL-80N which is an ethylene oxide propylene oxide alkoxylate of primary alcohol with a solubilization value of 1.022g/g
  • Tergitol N-10 which is an a trimethyl nonal ethoxylate with a solubilization value of 0.964g/g
  • Rexophos 25/97 which is a phosphated nonylphenol ethooxylate with a solubilization value of 0.951 g/g
  • the DLPLA polymer contains 65 %DL and 35 %PLG Weigh 0.02g triamcinolone into a glass vial
  • Aqueous phase Aqueous phase:
  • the SDS may be washed by continuously exchanging the solution mixture with DI water
  • Example 21 Biodegradable Microcapsule Containing Anti-proliferative Pharmaceutical a. Delayed 60 days b. Controlled release over 365 days Organic phase:
  • the DLPLA polymer contains 100%PGA
  • Aqueous phase Aqueous phase:
  • the SDS may be washed by continuously exchanging the solution mixture with DI water
  • Example 22 Dermal Filler Composition Containing anesthetic, Cortical Steroid and Antiproliferative Pharmaceutical a. Biodegradable microcapsule containing a cortical steroid delayed 30 days, controlled release over 120 days b. Biodegradable microcapsule containing an anti-pro liferative pharmaceutical delayed 60 days, controlled released over 365 days
  • composition mixture dry
  • Hyaluronic acid cross-linked 60% - 95%
  • Anti-inflammatory drug containing micro particles 5% - 20%
  • Antiproliferative drug containing micro particles 5% - 20%
  • Anesthetic drug (lidocaine hydrochloride) 0.1% - 5%
  • Example 23 Encapsulation of an anti-proliferative pharmaceutical a biodegradable acrylic acid copolymer
  • Aqueous phase is: o 75mL of 0.5% polyvinyl alcohol solution maintained at room temperature
  • the polyvinyl alcohol may be washed by continuously exchanging the solution mixture with fresh DI water
  • the other important characteristics of the gel slurries according to the one embodiment which determine their usefulness in various medical fields is the complex combination of their rheological properties. These properties include viscosity and its dependence on shear rate, the ratio between elastic and viscous properties in dynamic mode, relaxation behavior and some others which are discussed below in more detail.
  • the rheology of the products of the one embodiment can be controlled over very broad limits, essentially by two methods.
  • the rheological properties of each of the two phases forming the viscoelastic gel slurry are controlled in such a way that gives the desirable rheology for the final product.
  • the second such method of controlling the rheology of the gel slurry consists of selecting a proper ratio for two phases. But because these parameters, i.e. rheology of the two phases and their ratio determine some other important properties of the products of one embodiment, the best way to control the rheology should be selected ad hoc for each specific case.
  • the gels suitable for the use in the products according to the one embodiment can represent very many different kinds of rheological bodies varying from hard fragile gels to very soft deformable fluid- like gels.
  • a conventional gelatin gel the hardness and elasticity of the gel increases with increasing polymer concentration.
  • the rheological properties of a crosslinked gel are usually a function of several parameters such as crosslinking density, polymer concentration in the gel, composition of the solvent in which the crosslinked polymer is swollen. Gels with different rheological properties based on hyaluronan and hylan are described in the above noted U.S. Pat. Nos.
  • the rheological properties of the gel can be controlled, mainly, by changing the polymer concentration in the starting reaction mixture and the ratio of the polymer and the crosslinking agent, vinyl sulfone. These two parameters determine the equilibrium swelling ratio of the resulting gel and, hence, the polymer concentration in the final product and its rheological properties.
  • a substantial amount of solvent can be removed from a gel which had previously been allowed to swell to equilibrium, by mechanical compression of the gel.
  • the compression can be achieved by applying pressure to the gel in a closed vessel with a screen which is permeable to the solvent and impermeable to the gel.
  • the pressure can be applied to the gel directly by means of any suitable device or through a gas layer, conveniently through the air.
  • the other way of compressing the gel is by applying centrifugal force to the gel in a vessel which has at its bottom the above mentioned semipermeable membrane.
  • the compressibility of a polymeric gel slurry depends on many factors among which are the chemical nature of the gel, size of the gel particles, polymer concentration and the presence of a free solvent in the gel slurry.
  • Partial removal of the solvent from a gel slurry makes the slurry more coherent and substantially changes the rheological properties of the slurry.
  • the magnitude of the changes strongly depends on the degree of compression, hereinafter defined as the ratio of the initial volume of the slurry to the volume of the compressed material.
  • the achievable degree of compression i.e. compressibility of a gel slurry
  • the polymer concentration in the gel phase of the viscoelastic mixtures may vary over broad ranges depending on the desired properties of the mixtures which, in turn, are determined by the final use of the mixture. In general, however, the polymer concentration in the gel phase can be from 0.01 to 30%, preferably, from 0.05 to 20%. In the case of hylan and hyaluronan pure or mixed gels, the polymer concentration in the gel is preferably, in the range of 0.1 to 10%, and more preferably, from 0.15 to 5% when the swelling solvent is physiological saline solution (0.15M aqueous sodium chloride).
  • the choice of a soluble polymer or polymers for the second phase of the viscoelastic gel slurries is governed by many considerations determined by the final use of the product.
  • the polymer concentration in the soluble polymer phase may vary over broad limits depending on the desired properties of the final mixture and the properties of the gel phase. If the rheological properties of the viscoelastic gel slurry are of prime concern then the concentration of the soluble polymer may be chosen accordingly with due account taken of the chemical nature of the polymer, or polymers, and its molecular weight.
  • the polymer concentration in the soluble phase may be from 0.01% to 70%, preferably from 0.02 to 40%.
  • hylan or hyaluronan are used as the soluble polymers
  • their concentration may be in the range of 0.01 to 10%, preferably 0.02 to 5%.
  • other glycosaminoglycans such as chondroitin sulfate, dermatan sulfate, etc.
  • their concentration can be substantially higher because they have a much lower molecular weight.
  • the two phases forming the viscoelastic gel slurries according to one embodiment can be mixed together by any conventional means such as any type of stirrer or mixer.
  • the mixing should be long enough in order to achieve uniform distribution of the gel phase in the polymer solution.
  • the gel phase may already be a slurry obtained by disintegrating a gel by any conventional means such as pushing it through a mesh or a plate with openings under pressure, or by stirring at high speed with any suitable stirrer.
  • the viscoelastic mixed gel slurries can be prepared by mixing large pieces of gel with the polymer solution and subsequently disintegrating the mixture with formation of the viscoelastic slurry by any conventional means discussed above.
  • the gel slurry phase can be made of a gel swollen to equilibrium, and in this case there is no free solvent between the gel particles, or it may have some free solvent between gel particles. In the latter case this free solvent will dilute the polymer solution used as the second phase.
  • the third type of gel slurry used as the gel phase in the mixture is a compressed gel whose properties were discussed above. When a compressed gel slurry is mixed with a polymer solution in some cases the solvent from the solution phase will go into the gel phase and cause additional swelling of the gel phase to equilibrium when the thermodynamics of the components and their mixture allows this to occur.
  • composition of the viscoelastic mixed gel slurries can vary within broad limits.
  • the polymer solution in the mixture can constitute from 0.1 to 99.5%, preferably, from 0.5 to 99%, more preferably, from 1 to 95%, the rest being the gel phase.
  • the choice of the proper composition of the mixture depends on the properties and composition of the two components and is governed by the desirable properties of the slurry and its final use.
  • the viscoelastic gel mixtures according to one embodiment in addition to the two major components, namely, the polymeric gel slurry and the polymer solution, may contain many other components such as various physiologically active substances, including drugs, fillers such as micro crystalline cellulose, metallic powders, insoluble inorganic salts, dyes, surface active substances, oils, viscosity modifiers, stabilizers, etc., all depending upon the ultimate use of the products.
  • various physiologically active substances including drugs, fillers such as micro crystalline cellulose, metallic powders, insoluble inorganic salts, dyes, surface active substances, oils, viscosity modifiers, stabilizers, etc., all depending upon the ultimate use of the products.
  • the viscoelastic gel slurries represent, essentially, a continuous polymer solution matrix in which discrete viscoelastic gel particles of regular or irregular shape are uniformly distributed and behave Theologically as fluids, in other words, they exhibit certain viscosity, elasticity and plasticity.
  • compositional parameters of the slurry namely the polymer concentration in the gel and the solution phases, and the ratio between two phases
  • one may conveniently control the rheological properties of the slurry such as the viscosity at a steady flow, elasticity in dynamic mode, relaxation properties, ratio between viscous and elastic behavior, etc.
  • the other group of properties which are strongly affected by the compositional parameters of the viscoelastic gel slurries relates to diffusion of various substances into the slurry and from the slurry into the surrounding environment.
  • the diffusion processes are of great importance for some specific applications of the viscoelastic gel slurries in the medical field such as prevention of adhesion formation between tissues and drug delivery as is discussed below in more detail.
  • adhesion formation between tissues is one of the most common and extremely undesirable complications after almost any kind of surgery.
  • the mechanism of adhesion formation normally involves the formation of a fibrin clot which eventually transforms into scar tissue connecting two different tissues which normally should be separated.
  • the adhesion causes numerous undesirable symptoms such as discomfort or pain, and may in certain cases create a life threatening situation.
  • the adhesion formation requires another operation just to eliminate the adhesions, though there is no guarantee against the adhesion formation after re-operation.
  • One means of eliminating adhesion is to separate the tissues affected during surgery with some material which prevents diffusion of fibrinogen into the space between the tissues thus eliminating the formation of continuous fibrin clots in the space.
  • a biocompatible viscoelastic gel slurry can be successfully used as an adhesion preventing material.
  • the diffusion of low and high molecular weight substances in the case of plain gel slurries can easily occur between gel particles especially when the slurry mixes with body fluids and gel particles are separated from each other.
  • a viscoelastic mixed gel slurry according to one embodiment is implanted into the body, the polymer solution phase located between gel particles continues to restrict the diffusion even after dilution with body fluids thus preventing adhesion.
  • this effect would be more pronounced with an increase in polymer concentration of the polymer solution phase.
  • each of the phases of the slurry or both phases can be loaded with a drug or any other substance having physiological activity which will slowly diffuse from the viscoelastic slurry after its implantation into the body and the diffusion rate can be conveniently controlled by changing the compositional parameters of the slurries.
  • Components of the viscoelastic mixed gel slurries affect the behavior of living cells by slowing down their movement through the media and preventing their adhesion to various surfaces.
  • the degree of manifestation of these effects depends strongly on such factors as the composition of the two components of the mixture and their ratio, the nature of the surface and its interaction with the viscoelastic gel slurry, type of the cells, etc. But in any case this property of the viscoelastic gel slurries can be used for treatment of medical disorders where regulation of cell movement and attachment are of prime importance in cases such as cancer proliferation and metastasis.
  • biocompatible viscoelastic gel slurries include soft tissue augmentation, use of the material as a viscosurgical tool in opthalmology, otolaryngology and other fields, wound management, in orthopedics for the treatment of osteoarthritis, etc.
  • the following basic properties of the mixed gel slurries are utilized: biocompatibility, controlled viscoelasticity and diffusion characteristics, easily controlled residence time at the site of implantation, and easy handling of the material allowing, for example its injection through a small diameter needle. The following methods were used for characterization of the products obtained according to one embodiment.
  • the concentration of hylan or hyaluronan in solution was determined by hexuronic acid assay using the automated carbazole method (E. A. Balazs, et al, Analyt. Biochem. 12, 547-558, 1965).
  • the concentration of hylan or hyaluronan in the gel phase was determined by a modified hexuronic acid assay as described in Example 1 of U.S. Pat. No.4,582,865.
  • This material was then activated with perioxidate and then modified with an 18- amino acid peptide containing a cell attachment domain, Arg-Gly-Asp (RGD), to enhance cell attachment to the hydrogel.
  • RGD Arg-Gly-Asp
  • divinyl sulfone also cross-links hyaluronan, most likely via reaction with hydroxyl groups.
  • divinyl sulfone also cross-links hyaluronan, most likely reaction with hydroxyl groups.
  • the autocross-linked polymer (ACPTM, Fidia) is an internally esterified derivative of hyaluronan, with both inter- and intra-molecular bonds between the hydroxyl and carboxyl groups of hyaluronan.
  • ACPTM can be lyophilized to a white powder and hydrated to a transparent gel. This novel biomaterial has been used as a barrier to reduce post-operative Photo-cross Linking
  • a methacrylate derivative of hyaluronan was synthesized by the esterification of the hydroxyls with excess methacrylic anhydride, as described above for hyaluronan butyrate. This derivative was photocross-linked to form a stable hydrogel using ethyl eosin in l-vinyl-2-pyrrolidone and triethanolamine as an initiator under argon ion laser irradiation at 514 nm.
  • the use of in situ photopolymerization of an hyaluronan derivative which results in the formation of a cohesive gel enveloping the injured tissue, may provide isolation from surrounding organs and thus prevent the formation of adhesions.
  • a preliminary cell encapsulation study was successfully performed with islets of Langerhans to develop a bioartificial source of insulin. Glutaraldehyde cross linking
  • Hyaluronan strands extruded from cation-exchanged sodium hyaluronate (1.6 MDa) were cross-linked in glutaraldehyde aqueous solution, although the chemical nature of this process was not identified.
  • the strand surfaces were then remodeled by attachment of poly-D- and poly-L-lysine.
  • the polypeptide-resurfaced hyaluronan strands showed good biocompatibility and promoted cellular adhesion.
  • Intergel® (FeHA, LifeCore) is a hydrogel formulation of hyaluronan formed by chelation with ferric hydroxide. Similar cross-linking of yaluronan has been the basis of preparations using copper, zinc, calcium, barium, and other chelating metals. The reddish FeHA gel is in development for prevention of post- surgical adhesions. Carbodiimide cross linking
  • Incert® is a bioresorbable sponge (Anika Therapeutics) prepared by cross-linking hyaluronan with a biscarbodiimide in aqueous isopropanol. This procedure takes advantage of the otherwise undesirable propensity of carbodiimides to react with hyaluronan to form N-acylureas. In this application, the formation of two N- acylurea linkages provides a chemically stable and by-product-free cross-link. Because of the hydrophobic biscarbodiimides employed, Incert® adheres to tissues without the need for sutures and retains its efficacy even in the presence of blood. Recently, it was found to be effective at preventing post-operative adhesions in a rabbit fecal abrasion study.
  • a low-water content hyaluronan hydrogel film was made by cross-linking a hyaluronan (1.6 MDa) film with a water-soluble carbodiimide as a coupling agent in an aqueous mixture containing a water-miscible non-solvent of hyaluronan.
  • the highest degree of cross-linking that gave a low-water content hydrogel was achieved in 80% ethanol.
  • the cross-linking of hyaluronan films with a water-soluble carbodiimide in the presence of L-lysine methyl ester further prolonged the in vivo degradation of a hyaluronan film.
  • hydrogels have been prepared using bishydrazide, trishydrazide, and polyvalent hydrazide compounds as cross- linkers.
  • gels with physicochemical properties ranging from soft-pourable gels to more mechanically-rigid and brittle gels could be obtained.
  • HA-ADH can be cross-linked using commercially-available small molecule homobifunctional cross-linkers More recently, an in situ polymerization technique was developed by cross-linking HA-ADH with a macromolecular cross-linker, PEG-dialdehyde under physiological conditions.
  • Biocompatible and biodegradable hyaluronan hydrogel films with well-defined mechanical strength were obtained after the evaporation of solvent. Macromolecular drugs were released slowly from these hyaluronan hydrogel films, and these new materials accelerated re-epithelialization during wound healing.
  • Hylans are hydrogels or hydrosols formed by cross- linking hyaluronan-containing residual protein with formaldehyde in a basic solution.13 Soluble hylan is a high molecular weight form (8 - 23 MDa) of hyaluronan that exhibits enhanced rheological properties compared to hyaluronan. Hylan gels have greater elasticity and viscosity than soluble hylan materials, while still retaining the high biocompatibility of native hyaluronan. Hylans have been investigated in a number of medical applications. Multi-component reactions These are 3 to 4 component reactions known as (1) the Passerini reaction and (2) Ugi reactions.
  • an aqueous solution of hyaluronan is mixed with aqueous glutaraldehyde (or another water-soluble dialdehyde) and added to a known amount of a highly reactive isocyanide, e.g., cyclohexylisocyanide.
  • a highly reactive isocyanide e.g., cyclohexylisocyanide
  • the degree of cross-linking is controlled by the amount of aldehyde and diamine.
  • One example has to do with the Surfaces of polypropylene (PP) and polystyrene (PS) were activated with argon gas and ammonia gas plasmas to emanate the polymer surface. Emanated surfaces were then modified with succinic anhydride to give pendant carboxylic acid groups on the surface, which were then condensed with HA-ADH in the presence of a carbodiimide to give hydrophilic, non-adhesive, and lubricious plastic surfaces. Metal and glass surfaces can also be modified by surface activation followed by covalent chemical attachment of an appropriate hyaluronan derivative.
  • HA can be cross-linked at two locations: (1) the hydroxyl location and (2) the carboxyl location.
  • Drugs that have functional groups that favor reacting with hydroxyl and/or carboxyl could be conjugated on the HA molecule, and the HA molecule will act as a carrier of the drug.
  • Individual HA molecules could be grafted or attached covalently to a polymer chain that has pendant function groups which favor reacting with hydroxyl and/or carboxyl.
  • D. HA molecules can be grafted onto a liposome provided that their function groups favor reacting.
  • HA hydrogel Include cross-linked HA hydrogel, HA drug bioconjugate, HA-grafted copolymers, and HA liposomes
  • Esterified hyaluronan biomaterials have been prepared by alkylation of the tetra (n-butyl) ammonium salt of hyaluronan with an alkyl halide in dimethylformamide (DMF) solution.
  • DMF dimethylformamide
  • These hyaluronan esters can be extruded to produce membranes and fibers, lyophilized to obtain sponges, or processed by spray-drying, extraction, and evaporation to produce microspheres.
  • These polymers show good mechanical strength when dry, but the hydrated materials are less robust.
  • the degree of esterification influences the size of hydrophobic patches, which produces a polymer chain network that is more rigid and stable, and less susceptible to enzymatic degradation.
  • the chemical modification of the carboxylic functions of hyaluronan by carbodiimide compounds is generally performed in water at pH 4.75.
  • the sulfated hyaluronic acid HyalS3.5 was then immobilized onto plasma-processed polyethylene (PE) using a diamine polyethylene glycol derivative and a water-soluble carbodiimide.
  • PE polyethylene
  • HyalSx was converted to a photo labile azidophenylamino derivative and was photoimmobilized onto a poly(ethylene terephthalate) (PET) film.9
  • PET poly(ethylene terephthalate)
  • Hyaluronan butyrate is used as targeted drug-delivery system specifically to tumor cells.
  • Butyric acid is known to induce cell differentiation and to inhibit the growth of a variety of human tumors was coupled to hyaluronan via the reaction between butyric anhydride and the sym-collidinium salt of low molecular weight hyaluronan in DMF containing dimethy laminopyridine .
  • the anthracycline antibiotics adriamycin and daunomycin were coupled to hyaluronan via cyanogen bromide (CNBr) activation.
  • CNBr cyanogen bromide
  • This reaction scheme is commonly used to activate oligosaccharides to produce affinity matrices via a highly-reactive isourea intermediate.
  • the therapeutic agents appear to become attached via a urethane bond to one of the hydro xylic functions of the oligosaccharide or the glycosaminoglycan, but no spectroscopic verification was provided.
  • the harshness of the reaction conditions may compromise the integrity and biocompatibility of the hyaluronan.
  • Reactive bisaldehyde functionalities can be generated from the vicinal secondary alcohol functions on hyaluronan by oxidation with sodium peroxidase.
  • This chemistry is a standard method for chemical activation of glycoproteins for affinity immobilization or conversion to a fluorescent probe.
  • peroxidase-activated hyaluronan reductive coupling with primary amines can give cross-linking, attachment of peptides containing cell attachment domains, or immobilized materials.
  • the harsh oxidative treatment also introduces chain breaks and potentially immunogenic linkages into the hyaluronan biomaterial.
  • Reductive amination of the reducing end of hyaluronan has been employed to prepare affinity matrices, fluorophore-labeled materials, and hyaluronan- phospholipids for insertion into hyaluronan-liposomes.
  • low molecular weight hyaluronan was covalently attached to phosphatidyl- ethanolamine, and this conjugate has been employed for a protective "sugar decoration" on the surface of low density lipoprotein (LDL) particles.
  • LDL low density lipoprotein
  • the Materials can include
  • the reaction is a water in oil emulsion reaction
  • the X-linker mix must be used sooner than 24 hours after made up and kept at RT conditions
  • reaction temperature of 50C is too high to be kept for more than 1 hour.
  • the HA can be serially cross-linked to form a system with monophasic characteristics.
  • the forming a biocompatible cross-linked polymer as an IPN can be done by cross-linking a heteropolysaccharide to form a single cross-linked material; and performing one or more additional cross-linkings on the single cross-linked material to form a multiple cross-linked material, wherein the multiple cross-linked material has a core that lasts longer in a human body than the single cross-linked material.
  • the result is a material with a smooth continuum from slightly cross-linked to the core which is highly cross-linked.
  • the slightly cross-linked material enables the HA to be easily inserted into the human body with a small gauge syringe, but such slightly cross-linked material will not last long in the human body. However, the highly cross-linked material will remain longer in the human body so that the body augmentation does not need periodic touch-ups as is needed by conventional HA dermal fillers.
  • cross-link time resulting from the use of a stable, non-aqueous suspension of a delayed cross-linker according to the preferred embodiment may be controlled by varying any one or all of the following:
  • the type of molecular weight of the HA compound may be employed effectively to control the exact cross-linking time of the water- soluble solution. More particularly, suspensions of larger molecular weight HA cross-link more slowly than suspensions of low molecular weight acid.
  • the pH of the water soluble polymer solution prior to its cross-linking may be used to control cross-link time.
  • the pH of the water soluble polymer solution affects the solubility rate of the stable, non-aqueous suspension of a delayed cross-linker. Specifically, as the pH of the water soluble polymer solution increases, the solubility rate of the cross-linker suspension increases if the suspension contains a majority of HA particles, whereas the solubility rate of the cross-linker suspension decreases if the suspension contains a majority of borax particles.
  • the solubility rate of the cross-linker suspension decreases if the suspension contains a majority of boric acid particles, whereas the solubility rate of the cross-linker suspension increases if the suspension contains a majority of HA particles.
  • Both the concentration (i.e., loading) of the stable, non-aqueous suspension of a delayed HA cross-linker in the water soluble polymer solution and the content of the cross-linker suspension affect the cross-link time of a water soluble polymer solution similarly.
  • the concentration of the suspension of delayed HA cross-linker in the water-soluble polymer solution or the content of the cross-linker suspension increase, the cross-link time of the water soluble polymer solution decreases.
  • the concentration of the suspension of the delayed boron cross-linker in the water soluble polymer solution and the content of the cross- linker suspension decrease, the cross-link time of the water soluble polymer solution increases.
  • Temperature may be used to alter the cross-link time of a water soluble polymer solution. As the temperature of the water soluble polymer solution increases, its cross-link time decreases. Conversely, as the temperature of the water soluble polymer solution decreases, its cross-link time increases. Furthermore, the cross-link time of a water-soluble polymer may be increased or decreased depending upon the clay type utilized in the formulation of the stable, non-aqueous suspension of a delayed HA cross-linker.
  • materials such as polymeric microspheres, polymer micelles, soluble polymers and hydrogel-type materials can be used for providing protection for pharmaceuticals against biochemical degradation, and thus have shown great potential for use in biomedical applications, particularly as components of drug delivery devices.
  • biomedical polymers e.g., polymers for use under physiological conditions
  • polymeric materials must be compatible with the biological milieu in which they will be used, which often means that they show certain characteristics of hydrophilicity. They also have to demonstrate adequate biodegradability (i.e., they degrade to low molecular weight species.
  • the polymer fragments are in turn metabolized in the body or excreted, leaving no trace).
  • Biodegradability is typically accomplished by synthesizing or using polymers that have hydrolytically unstable linkages in the backbone.
  • the most common chemical functional groups with this characteristic are esters, anhydrides, orthoesters, and amides. Chemical hydrolysis of the hydrolytically unstable backbone is the prevailing mechanism for the degradation of the polymer.
  • Biodegradable polymers can be either natural or synthetic. Synthetic polymers commonly used in medical applications and biomedical research include polyethyleneglycol (pharmacokinetics and immune response modifier), polyvinyl alcohol (drug carrier), and poly(hydroxypropylmetacrylamide) (drug carrier). In addition, natural polymers are also used in biomedical applications.
  • dextran, hydroxyethylstarch, albumin and partially hydrolyzed proteins find use in applications ranging from plasma substitute, to radiopharmaceutical to parenteral nutrition.
  • synthetic polymers may offer greater advantages than natural materials in that they can be tailored to give a wider range of properties and more predictable lot-to-lot uniformity than can materials from natural sources.
  • the linker is a dicarboxylic acid with at least three atoms between the carbonyls and contains a heteroatom alpha to the carbonyl forming the ester, the release half-life is less than about 10 hours; when Linker is a dicarboxylic acid with at least three atoms between the carbonyls with no heteroatom alpha to the carbonyl forming the ester, the release half-life is more than about 100 hours; wherein when Linker is a dicarboxylic acid with two atoms between the carbonyls and Tether contains a nitrogen with a reactive hydrogen, the release half-life of the HA is from about 0.1 hours to about 20 hours; wherein the release half-life being measured in
  • the polyal is an acetal. In other embodiments, the polyal is a ketal. In some embodiments, the acetal is PHF. In some embodiments, Ri is H. In other embodiments, Ri is CH3. In some embodiments, R2 is -CH(Y)-C(0)-, wherein Y is one of the side chains of the naturally occurring amino acids. In some embodiments, R2 is an aryl group. In some embodiments, R2 is an heteroaryl group. In other embodiments, R2 is an aliphatic ring. In some embodiments, R2 is an aliphatic chain. In some embodiments, R2 is a heterocyclic aliphatic ring. In some embodiments, Ri and R2 when taken together with nitrogen to which they are attached form a ring. Other embodiments are known to those skilled in the art. For example, some embodiments are discussed in US2010/036413, the content of which is incorporated by reference.
  • FIG. 1 shows an exemplary system to serially produce multiply cross-linked HA.
  • HA material P-15 and sodium hydroxide P-16 is provided to a gate and measurement unit PI 4.
  • the output is provided to a mixer PI 7.
  • a cross-linker source E9 is provided to a reactor 1-7 whose output is stored at a tank P21. The stored cross-linked HA can then be atomized.
  • FIG. 2 shows another exemplary system to serially produce multiply cross-linked HA.
  • HA and sodium hydroxide is provided to a reactor that receives a plurality of cross-linker sources such as PVS1, PVS2, and PVS3 sources.
  • the reactor generated serially and multiply cross-linked HA is then cleaned at a chamber to remove residuals and to change pH to about 7.4.
  • the chamber receives distilled water and PBS at a pH of about 7.4.
  • the cleaned output is then sent to a final assembly and packaging station.
  • FIG. 3 shows an exemplary diagram of the resulting multiply cross-linked HA.
  • the composition includes a first portion 300 of a first polymer with lightly cross- linking extensions or arms; a second portion 310 of polymer with a first serially cross-linked center overlapping the first portion and one or more lightly cross-linked extensions adjacent the serially cross-linked center; and a third portion 320of polymer with a second serially cross- linked region 350 overlapping the second portion and one or more lightly cross-linked extensions adjacent the serially cross-linked center; wherein the lightly cross-linked extensions enable the composition to be injected through a small gauge needle and the second serially cross-linked center is resistant to absorbtion by biological processes.
  • the region 350 can be multiply cross- linked for biodegradation resistance.
  • the polymer can be one of: collagens, hyaluronic acids, celluloses, proteins, saccharides, an extracellular matrix of a biological system.
  • a biocompatible cross-linked IPN polymer can be done by cross- linking a heteropolysaccharide to form a first cross-linked material; and by performing one or more additional cross-linking of the first cross-linked material to form a multiple cross-linked material.
  • the result monophasic HA can be used for augmenting soft tissue with the biocompatible cross-linked polymer.
  • semi-IPN can also be obtained by the polymerization of a monomer in the presence of a crosslinking agent and in the presence of the natural acidic polysaccharide or a semisynthetic ester-type derivative thereof.
  • the HA composition percentage is varied from 75% to 99% of the total composition while the cross linker percentage is varied between 1 and 25% as follows:
  • the material is soft, but less resistant to biodegration. As more cross-linker is introduced, the material becomes more hardened and lasts longer.
  • the multiple serially cross-linking processes provide advantages of being soft to the touch, yet long lasting.
  • the varying mechanical/physical properties that constantly becomes softer while remaining tough radiating out from the IPN makes the polymer tough and at the same time compliant with its surrounding for better biocompatibility and feels more natural to the touch.
  • the IPN is an intimate combination of two or more polymer systems, both in network form, at least one of which is synthesized or cross-linked in the immediate presence of the other.
  • IPN IPPN
  • the multiply cross linking process is akin to a discrete or digital process where the HA is first cross-linked, then the result is cross-linked a second time, then third cross-linked is done, thus forming serial cross-linking additions.
  • This discrete or digital process is in contrast to the conventional continuous process.
  • the IPN center can be where ever relative aqueous front exists. It should be mentioned that for the purpose of HA longevity, the more hydrophobic a cross linker is the better because hydrolysis is not favored. Sterically hindered cross linker is also preferred for the same reason mentioned. However, hydrophobicity in this case will make the HA polymer less biocompatible and will likely illicit unwanted foreign body reactions.
  • the type of cross linker used for any part of the process will also make a difference in longevity, biocompatibility and physical properties. Application requirement will dictate the ideal polymer composition that gives the balance of properties.
  • the HA can be used as facial fillers, dermal fillers, butt fillers, breast fillers, and other body part fillers.
  • the implants of the present invention further can be instilled, before or after implantation, with indicated medicines and other chemical or diagnostic agents.
  • Such agents include, but are not limited to, antibiotics, chemotherapies, other cancer therapies, brachy-therapeutic material for local radiation effect, x-ray opaque or metallic material for identification of the area, hemostatic material for control of bleeding, growth factor hormones, immune system factors, gene therapies, biochemical indicators or vectors, and other types of therapeutic or diagnostic materials which may enhance the treatment of the patient.
  • Advantages of one IPN embodiment can include one or more of the following.
  • a natural feel is achieved through viscoelastic harmony of properties between the existing tissue and the implant. This can be done by manipulating the viscous component of the implant through flow properties by way of the particle size and particle size distribution ratios.
  • the elastic component is intrinsic within the material tertiary structure (molecular weight and steric hindrance) and cross linking densities.
  • the interpenetrating polymer network hydrogels have a number of desirable properties. These properties include high tensile strength with high water content, making the interpenetrating polymer network hydrogels excellent for use in dermal filling applications.

Abstract

La présente invention concerne des systèmes et un procédé permettant de former un polymère réticulé biocompatible comportant un réseau polymère interpénétrant (IPN), par la réticulation d'un hétéropolysaccharide pour former une substance monoréticulée et la mise en œuvre d'une ou de plusieurs réticulations supplémentaires sur la substance monoréticulée pour former une substance multiréticulée. La substance multiréticulée comporte une ou plusieurs régions IPN non biodégradables dans un corps humain par rapport à la substance monoréticulée et une ou plusieurs extensions monoréticulées rayonnant à partir de l'IPN, la combinaison de l'IPN et de l'extension conférant une résistance à la biodégradation et une douceur au toucher et facilitant l'insertion dans le corps humain.
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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2017036597A1 (fr) * 2015-09-01 2017-03-09 Merz Pharma Gmbh & Co. Kgaa Procédé de fabrication d'un gel polymère réticulé et dispositif pour la mise en œuvre du procédé
US11400182B2 (en) 2019-11-22 2022-08-02 Gcs Co., Ltd. Injectable formulation containing a poly l lactic acid filler and a hyaluronic acid filler conjugate and a method for preparing the same
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EP3947546B1 (fr) * 2019-04-05 2023-04-19 Fina Technology, Inc. Polyenes pour des compositions durcissables à base de caoutchoucs liquids
US11642415B2 (en) 2017-03-22 2023-05-09 Ascendis Pharma A/S Hydrogel cross-linked hyaluronic acid prodrug compositions and methods
US11865225B2 (en) 2019-11-22 2024-01-09 Gcs Co., Ltd. Sustained release injectable formulation containing a poly L lactic acid filler and a hyaluronic acid filler conjugate and a method for preparing the same

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* Cited by examiner, † Cited by third party
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CN116019978B (zh) * 2023-02-09 2024-01-23 云南贝泰妮生物科技集团股份有限公司 一种微交联透明质酸钠-重组胶原蛋白复合凝胶及其制备方法和应用

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6224893B1 (en) * 1997-04-11 2001-05-01 Massachusetts Institute Of Technology Semi-interpenetrating or interpenetrating polymer networks for drug delivery and tissue engineering
US6281341B1 (en) * 1997-04-30 2001-08-28 Biomm, Inc. & University Of Miami Hetero-polysaccharide conjugate and methods of making and using the same
EP1167405A2 (fr) * 2000-05-02 2002-01-02 Dow Corning Corporation Polymère à réseaux interpénétrés à base de dendrimères
US20070026518A1 (en) * 2005-03-29 2007-02-01 The Regents Of The University Of California Controlling stem cell destiny with tunable matrices
US20100256671A1 (en) * 2009-04-07 2010-10-07 Biomedica Management Corporation Tissue sealant for use in noncompressible hemorrhage

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
IT1260154B (it) * 1992-07-03 1996-03-28 Lanfranco Callegaro Acido ialuronico e suoi derivati in polimeri interpenetranti (ipn)
US5827937A (en) * 1995-07-17 1998-10-27 Q Med Ab Polysaccharide gel composition
GB9902412D0 (en) * 1999-02-03 1999-03-24 Fermentech Med Ltd Process
GB9902652D0 (en) * 1999-02-05 1999-03-31 Fermentech Med Ltd Process
DE102006013594A1 (de) * 2006-03-22 2007-09-27 Biopolymer Gmbh & Co. Kg Quervernetzte Gele von Hyaluronsäure und deren Verwendung

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6224893B1 (en) * 1997-04-11 2001-05-01 Massachusetts Institute Of Technology Semi-interpenetrating or interpenetrating polymer networks for drug delivery and tissue engineering
US6281341B1 (en) * 1997-04-30 2001-08-28 Biomm, Inc. & University Of Miami Hetero-polysaccharide conjugate and methods of making and using the same
EP1167405A2 (fr) * 2000-05-02 2002-01-02 Dow Corning Corporation Polymère à réseaux interpénétrés à base de dendrimères
US20070026518A1 (en) * 2005-03-29 2007-02-01 The Regents Of The University Of California Controlling stem cell destiny with tunable matrices
US20100256671A1 (en) * 2009-04-07 2010-10-07 Biomedica Management Corporation Tissue sealant for use in noncompressible hemorrhage

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of EP2776077A4 *

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2017036597A1 (fr) * 2015-09-01 2017-03-09 Merz Pharma Gmbh & Co. Kgaa Procédé de fabrication d'un gel polymère réticulé et dispositif pour la mise en œuvre du procédé
US11642415B2 (en) 2017-03-22 2023-05-09 Ascendis Pharma A/S Hydrogel cross-linked hyaluronic acid prodrug compositions and methods
EP3947546B1 (fr) * 2019-04-05 2023-04-19 Fina Technology, Inc. Polyenes pour des compositions durcissables à base de caoutchoucs liquids
US11400182B2 (en) 2019-11-22 2022-08-02 Gcs Co., Ltd. Injectable formulation containing a poly l lactic acid filler and a hyaluronic acid filler conjugate and a method for preparing the same
US11865225B2 (en) 2019-11-22 2024-01-09 Gcs Co., Ltd. Sustained release injectable formulation containing a poly L lactic acid filler and a hyaluronic acid filler conjugate and a method for preparing the same
CN115651218A (zh) * 2022-08-25 2023-01-31 四川大学 一种用于制备可注射多酚-大分子粘附性水凝胶的方法

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CN103917256A (zh) 2014-07-09
JP2014521492A (ja) 2014-08-28
EP2776077A4 (fr) 2014-12-17
KR20140059238A (ko) 2014-05-15
KR20150111372A (ko) 2015-10-05
EP2776077A1 (fr) 2014-09-17

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