CN108379112A - Dermal filler compositions for fine line treatment - Google Patents

Dermal filler compositions for fine line treatment Download PDF

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CN108379112A
CN108379112A CN201810249654.6A CN201810249654A CN108379112A CN 108379112 A CN108379112 A CN 108379112A CN 201810249654 A CN201810249654 A CN 201810249654A CN 108379112 A CN108379112 A CN 108379112A
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composition
gel
skin
ha
months
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CN201810249654.6A
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G·N·恩吉康
俞小杰
刘福田
萨米特·包利华
N·J·马内西斯
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阿勒根公司
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Priority to US61/534,780 priority
Priority to PCT/US2012/052125 priority patent/WO2013028904A2/en
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Priority to CN201280055657.7A priority patent/CN104105474B/en
Publication of CN108379112A publication Critical patent/CN108379112A/en

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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS 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
    • A61L27/00Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/14Macromolecular materials
    • A61L27/20Polysaccharides
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K8/00Cosmetics or similar toilet preparations
    • A61K8/18Cosmetics or similar toilet preparations characterised by the composition
    • A61K8/30Cosmetics or similar toilet preparations characterised by the composition containing organic compounds
    • A61K8/67Vitamins
    • A61K8/676Ascorbic acid, i.e. vitamin C
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K8/00Cosmetics or similar toilet preparations
    • A61K8/18Cosmetics or similar toilet preparations characterised by the composition
    • A61K8/72Cosmetics or similar toilet preparations characterised by the composition containing organic macromolecular compounds
    • A61K8/73Polysaccharides
    • A61K8/735Mucopolysaccharides, e.g. hyaluronic acid; Derivatives thereof
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILET PREPARATIONS
    • A61Q19/00Preparations for care of the skin
    • A61Q19/08Anti-ageing preparations
    • CCHEMISTRY; METALLURGY
    • C08ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08BPOLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C08B37/006Heteroglycans, 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
    • C08B37/0063Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan
    • C08B37/0072Hyaluronic acid, i.e. HA or hyaluronan; Derivatives thereof, e.g. crosslinked hyaluronic acid (hylan) or hyaluronates
    • CCHEMISTRY; METALLURGY
    • C08ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08JWORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G
    • C08J3/00Processes of treating or compounding macromolecular substances
    • C08J3/24Crosslinking, e.g. vulcanising, of macromolecules
    • CCHEMISTRY; METALLURGY
    • C08ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08LCOMPOSITIONS OF MACROMOLECULAR COMPOUNDS
    • C08L5/00Compositions of polysaccharides or of their derivatives not provided for in groups C08L1/00 or C08L3/00
    • C08L5/08Chitin; Chondroitin sulfate; Hyaluronic acid; Derivatives thereof
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K2800/00Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/80Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
    • A61K2800/91Injection
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS 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
    • A61L2400/00Materials characterised by their function or physical properties
    • A61L2400/06Flowable or injectable implant compositions
    • CCHEMISTRY; METALLURGY
    • C08ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08JWORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G
    • C08J2305/00Characterised by the use of polysaccharides or of their derivatives not provided for in groups C08J2301/00 or C08J2303/00
    • C08J2305/08Chitin; Chondroitin sulfate; Hyaluronic acid; Derivatives thereof
    • CCHEMISTRY; METALLURGY
    • C08ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08JWORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G
    • C08J3/00Processes of treating or compounding macromolecular substances
    • C08J3/02Making solutions, dispersions or lattices by other methods than by solution, emulsion or suspension polymerisation techniques
    • C08J3/03Making solutions, dispersions or lattices by other methods than by solution, emulsion or suspension polymerisation techniques in aqueous media
    • C08J3/075Macromolecular gels

Abstract

The present invention provides highly injectable, long-lasting hyaluronic acid-based hydrogel dermal filler compositions which are particularly advantageous for correction of fine lines in the face.

Description

用于细纹治疗的真皮填充剂组合物 Dermal filler composition for the treatment of fine lines

[0001] 本申请是申请日为2012年9月13日、申请号为“201280055657.7”、发明名称为“用于细纹治疗的真皮填充剂组合物”的发明专利申请的分案申请。 [0001] This application is filed September 13, 2012, application number "201280055657.7", entitled "lines for treating dermal filler composition" Patent Application invention divisional application.

[0002] 相关申请案 [0002] Related Applications

[0003] 本申请要求2011年9月14日提交的美国临时专利申请No. 61 /534,780的优先权和利益,并且是2012年8月23日提交的美国专利申请序列号13/593,313的部分继续申请,后者是2012年6月1日提交的美国专利申请序列号13/486,754的部分继续申请,其要求了2011年6月3日提交的美国临时专利申请No. 61 /493,309的优先权和利益,这些申请案中每一个的全部公开内容通过这种特定引用整体并入本文。 [0003] This application claims priority to US Provisional Patent Sept. 14, 2011 and filed Application No. 61 benefit / 534,780, and US Patent No. 2012, filed August 23, Application Serial No. 13 / 593,313 to continuation application, which is part of US Patent application serial No. 13 / 486,754 of June 1, 2012 filed a continuation application, which claims priority to US provisional Patent June 3, 2011 filed No. 61 / 493,309 priority to and benefit of each of these applications the entire disclosure of which is incorporated herein by this specific reference.

[0004] 背景 [0004] BACKGROUND

[0005] 本发明通常涉及真皮填充剂组合物,并且更具体地涉及对治疗皮肤上的细纹有效的可注射真皮填充剂组合物。 [0005] The present invention generally relates to a dermal filler composition, and more particularly relates to the treatment of fine lines on the skin is effective injectable dermal filler composition.

[0006] 皮肤老化是一个渐进现象,随着时间推移而发生并且受生活方式因素影响,例如饮酒、吸烟和日照。 [0006] Skin aging is a gradual phenomenon, which occurs over time and is influenced by lifestyle factors such as alcohol consumption, smoking and sunshine. 可用萎缩、松弛和肥胖表征面部皮肤的老化。 Available atrophy, relaxation and obesity characterize aging facial skin. 萎缩对应于皮肤组织厚度的大大减小。 Atrophy of skin thickness corresponding to the tissue is greatly reduced. 皮下组织松弛导致皮肤过多和下垂并且导致出现脸颊和眼睑下垂。 Relaxation results in excessive skin and subcutaneous tissue and cause sagging cheeks and ptosis. 肥胖指因为面部和颈部底部肿胀而增加多余重量。 Obesity refers to a bottom swelling of the face and neck since the increased excess weight. 这些变化通常与干燥、失去弹性和肌理粗糙相关。 These changes usually associated with dry, rough-related loss of elasticity and texture.

[0007] 透明质酸(HA),也称为透明质烷,是广泛分布于整个人体的结缔、上皮和神经组织的非硫酸化糖胺聚糖。 [0007] Hyaluronic acid (HA), also known as hyaluronan, is widely distributed in the entire non-sulfated glycosaminoglycan human connective, epithelial and neural tissues. 透明质酸在不同皮肤层中都很丰富,其中透明质酸具有多种功能,例如确保良好的水合作用,帮助细胞外基质的构成,起填充材料的作用和参与组织修复机制。 Hyaluronic acid is very rich in different layers of the skin, wherein the hyaluronic acid has a variety of functions, such as ensuring good hydration, helps constitute the extracellular matrix, acts as filler material and participate in tissue repair mechanisms. 然而,透明质酸、胶原蛋白、弹性蛋白和皮肤中存在的其它母体聚合物的量随年龄减少。 However, hyaluronic acid, collagen, elastin, and the amount of other matrix polymer present in the skin decrease with age. 例如,反复暴露于(例如)来自太阳的紫外光引起真皮细胞减少其透明质烷的生成以及增加了透明质烷的降解速率。 For example, repeated exposure to (e.g.) the ultraviolet light from the sun which causes dermal cells reduce the production of hyaluronan and increases the rate of degradation of hyaluronan. 这种材料损失导致各种皮肤状况,例如起皱纹、凹窝、水分流失和有助于老化出现的其它不良状况。 This loss of materials lead to various skin conditions, such as wrinkles, dimples, moisture loss and other adverse conditions contribute to aging appear.

[0008] 可注射真皮填充剂已经成功用于治疗老化皮肤。 [0008] The injectable dermal fillers have been used successfully to treat aging skin. 填充剂可代替失去的内源母体聚合物,或增强/促进现有母体聚合物的功能,以便治疗这些皮肤状况。 The filler can replace the missing endogenous matrix polymer, or enhancement / promotion of matrix polymer prior to the treatment of these skin conditions. 基于透明质酸的真皮填充剂已经变得越来越流行,因为透明质酸是在整个人体自然存在的物质。 Hyaluronic acid-based dermal fillers have become increasingly popular because hyaluronic acid is a substance naturally present throughout the body. 这些填充剂通常耐受良好、非永久性,并且是对多种皮肤状况的相当低风险的治疗。 These fillers are usually well-tolerated, non-permanent, and the treatment of various skin conditions is fairly low risk.

[0009] 廷德尔效应(Tyndall effect)是在施用了基于透明质酸(HA)的真皮填充剂的一些患者中出现的不良事件。 [0009] Tyndall effect (Tyndall effect) is administered adverse events in some patients with hyaluronic acid (HA) dermal fillers based. 廷德尔效应特征在于,在已经注射了真皮填充剂的皮肤部位出现蓝色变色,这表示通过半透明表皮看到可见透明质酸。 Characterized in that the Tyndall effect, the skin site had been injected in the dermal filler is a blue color, which indicates that the hyaluronic acid see visible through the translucent skin. 临床报道表明,填充剂施用技术和皮肤性质可影响这种不良事件的表现。 Clinical reports indicate that the filler application technique and the nature of the skin can affect the performance of such adverse events. 高硬度和高弹性的填充剂成功用于修正面部上如鼻唇沟、脸颊和下巴等区域,无需对面部变色有任何担心,因为材料注射在中部和深部真皮区中。 High elasticity and high hardness fillers such as nasolabial folds successfully used, cheeks and chin area correction portion, without any fear of discoloration of the face, because the material is injected in the middle and deep dermis region. 然而,当使用这些填充材料修正浅表、细小皱纹,例如泪沟、眉间纹、眼角纹、笑纹或前额,或在真皮上部区域中太浅表地错误应用时,常常观察到皮肤的浅蓝色变色。 However, when these correction superficial filler, fine wrinkles, tears ditch e.g., frown lines, eye pattern, laugh lines or forehead, or dermis shallow upper region table to application errors, frequently observed blue skin discoloration. 这种现象, 被认为是廷德尔效应的结果,引起应用部位半永久性变色,并且有时候只有在施用了降解填充材料的透明质酸酶后消失。 This phenomenon is considered to be the result of the Tyndall effect, causing the application site semi-permanent color, and sometimes only after the administration of hyaluronidase degradation filler material disappears. 因此,廷德尔效应在治疗浅表细小皱纹的患者中更常见。 Therefore, the Tyndall effect is more common in patients in the treatment of superficial fine wrinkles. 只要凝胶维持在皮肤中,廷德尔效应表现延长,通常几个月,是患者中主要关注点的一个原因。 As long as the gel to maintain in the skin, Tyndall effect the performance of the extension, usually a few months, it is a major cause of patient concerns.

[0010]已经特别配制了基于HA的真皮填充凝胶以治疗泪沟、前额、眼角纹、眉间纹等周围发现的“细小皱纹”。 [0010] in particular has been formulated based dermal filler HA gel to treat peripheral tear trough, forehead, eye pattern, frown lines and the like found in "fine wrinkles." 市场上可买到的HA“细纹”凝胶包括Juv6derm Refine (G'〜67Pa;G”/G' 〜O · 59,HA 浓度18mg/ml)、Belotero Soft (G' 〜28Pa; G” /G' 〜1 · I,HA 浓度20mg/ml)、 Eme;rvelTouch(G'〜56Pa;G”/G'〜0·64,HA浓度20mg/ml)、StylageS(G'〜192Pa;G”/G'〜 0· 20,HA浓度16mg/ml)、Teosyal First Lines (G'59Pa;G”/G' 〜0· 53,HA浓度20mg/ml)、 Restylane Touch(G' 〜489Pa;G”/G' 〜0.24,HA浓度18mg/ml)。 Commercially available HA "lines" gel comprising Juv6derm Refine (G'~67Pa; G "/ G '~O · 59, HA concentration of 18mg / ml), Belotero Soft (G' ~28Pa; G" / G '~1 · I, HA concentration of 20mg / ml), Eme; rvelTouch (G'~56Pa; G "/ G'~0 · 64, HA concentration of 20mg / ml), StylageS (G'~192Pa; G" / G'~ 0 · 20, HA concentration of 16mg / ml), Teosyal First Lines (G'59Pa; G "/ G '~0 · 53, HA concentration of 20mg / ml), Restylane Touch (G' ~489Pa; G" / G '~0.24, HA concentration of 18mg / ml). 虽然(例如)通过使线性HA链与少量交联剂轻微交联和/或减小这些凝胶的最终HA浓度,将这些凝胶配制成具有低弹性模量,但是大部分市场上可买到的“细纹”凝胶在一些患者中仍显示出廷德尔效应,特别是在浅表注射时,例如在小于约Imm的深度。 Although the (e.g.), and lightly crosslinked by a crosslinking agent and a small amount of linear chains HA / or reducing the final concentration of HA in these gels, these gels formulated to have a low modulus of elasticity, but the majority of commercially available the "lines" gel still showed Tyndall effect in some patients, particularly at shallow injection, for example, at a depth of less than about Imm.

[0011] 在浅表皱纹的治疗中可采用基于胶原蛋白的凝胶并且似乎不会引起廷德尔效应。 [0011] Tyndall effect can be based on collagen gels and does not appear to cause the treatment of superficial wrinkles. 基于胶原蛋白的凝胶未受到高度青睐,因为它们在皮肤中的持续时间相对较差并且需要在个体中预先测试。 Collagen-based gels is not highly favored because they duration in the skin and need to pre-test is relatively poor in an individual. _RadiesseK (轻基磷灰石钙)是皮下、可注射植入物,其主要组分是合成羟基磷灰石钙,而非透明质酸。 _RadiesseK (light calcium hydroxyapatite) subcutaneously, an injectable implant, the main component of synthetic calcium hydroxyapatite, hyaluronic acid instead. 与基于透明质酸的真皮填充剂不同,羟基磷灰石钙不透明,因此避免了廷德尔效应的并发症。 And calcium hydroxyapatite opaque hyaluronic acid-based dermal fillers different, thus avoiding the complications of the Tyndall effect. 然而,如果放置太浅,这种填充剂可被视为直接位于皮肤下的白色物质。 However, if placed too shallow, the filler can be considered white material located directly under the skin. 此外,与基于透明质酸的填充剂相比,Radiessek需要较长针头来注射并且通常不推荐用于眼部区域。 Furthermore, compared to hyaluronic acid-based fillers, Radiessek to require a longer injection needle and generally not recommended for the eye area.

[0012] 将需要提供不会表现出归因于廷德尔效应的浅蓝色变色的基于透明质酸的可注射真皮填充剂。 [0012] A need to provide not exhibit a Tyndall effect attributed to bluish discoloration of hyaluronic acid-based dermal fillers injectable.

[0013] 概述 [0013] Overview

[0014] 本发明描述了制备可在上部真皮中施用,不产生皮肤的任何浅蓝色变色,或至少不显著或不明显的浅蓝色变色的基于HA的真皮填充剂的组合物和配制方法。 [0014] The present invention describes the preparation can be administered in an upper dermis, bluish discoloration of the skin does not produce any or at least no significant or insignificant bluish discoloration of HA dermal fillers and formulation of compositions based on . 进一步地,已经发现目前描述的许多本发明的填充凝胶在体内持续时间明显比当前市场上可买到的凝胶更长。 Further, it has been found that many gel-filled presently described invention significantly longer in vivo duration than the current commercially available gels. 在本发明的一些方面,提供了对增强皮肤外观有用的光学透明的真皮填充剂,其增加了体积和丰满度,并且减少了甚至细小皱纹的出现,没有“廷德尔效应”。 In some aspects of the present invention, it provides a useful optically transparent dermal fillers for enhancing the appearance of skin, which increases the size and fullness, and is reduced even small wrinkles, no "Tyndall effect." 本组合物可引入皮肤的细纹,甚至薄皮区域中并且相当浅表,不会引起与许多常规光学透明的真皮填充剂相关的负面蓝色变色。 The present composition may be introduced into the fine lines of the skin, and even quite superficial thin skin region without causing many conventional dermal fillers optically clear negative correlation blue color.

[0015] 更具体地,在本发明的一个方面,提供了长效治疗性真皮填充剂组合物,其通常包含生物相容性聚合物,例如交联透明质酸组分和与透明质酸组分合并的添加剂。 [0015] More specifically, in one aspect of the present invention, there is provided a long-term therapeutic dermal filler composition, which typically comprises a biocompatible polymer, such as cross-linked hyaluronic acid and a hyaluronic acid component group were combined additives.

[0016] 在一个实施方案中,聚合物为多糖,例如透明质酸。 [0016] In one embodiment, the polymer is a polysaccharide, such as hyaluronic acid. 透明质酸包括交联组分并且可进一步包括非交联组分。 Comprising hyaluronic acid and a crosslinking component may further comprise a non-crosslinking component. 添加剂可包括维生素,例如维生素C (例如,稳定形式的维生素C)或维生素C衍生物(例如,L-抗坏血酸2-葡萄糖苷(AA2G)、抗坏血酸3-氨丙基磷酸酯(维生食物(Vitagen))或抗坏血酸磷酸钠(AA2P))。 Additives can include vitamins such as vitamin C (e.g., stable form of vitamin C) or a vitamin C derivative (e.g., L- ascorbic acid 2-glucoside (of AA2G), 3-aminopropyl ascorbyl phosphate (Vitagen (Vitagen )) or sodium ascorbyl phosphate (AA2P)).

[0017] 在本发明的一个方面,添加剂是维生素衍生物,其通过适合反应过程,例如醚化、 酰胺化或酯化与聚合物共价结合。 [0017] In one aspect of the invention, the additive is a vitamin derivative, by reaction of suitable processes, for example etherification, esterification or amidation is covalently bound to the polymer.

[0018] 在本发明的一个广泛方面,提供了真皮填充剂组合物,所述组合物包含与交联组分交联的透明质酸组分和除所述交联组分以外的添加剂。 [0018] In one broad aspect of the present invention, there is provided a dermal filler composition, the composition comprising hyaluronic acid with a crosslinking component and a crosslinking component additives other than the crosslinking component. 透明质酸组分可与添加剂化学结合。 Hyaluronic acid component can be combined with chemical additives. 进一步地,当施用到患者的真皮区内时,相对于除没有所述添加剂外,大体上相同的组合物,所述组合物表现出廷德尔效应减小。 Further, when administered to a patient dermal region, without addition of the additive with respect to an outer, substantially the same composition, said composition exhibits reduced Tyndall effect. 所述组合物可进一步包含其它添加剂,例如麻醉剂(例如利多卡因)。 The composition may further comprise other additives, for example, anesthetic (such as lidocaine). 在一个实施方案中,添加剂是维生素C衍生物,例如AA2G。 In one embodiment, the additive is a vitamin C derivative, e.g. AA2G. 在另一实施方案中,添加剂是维生食物。 In another embodiment, the additive is Vitagen.

[0019] 在一个实施方案中,透明质酸组分与添加剂化学结合,结合度介于约3mol %和约40mol之间,例如介于约3mol %和约IOmol %之间。 [0019] In one embodiment, the hyaluronic acid component and the additive chemically bound, binding of between about 3mol% and about 40 mol, for example between about 3mol% and about IOmol%.

[0020] 所述组合物可能大体上光学透明。 [0020] The composition may be substantially optically transparent. 所述组合物的G'值通常介于约40Pa和约IOOPa 之间,例如不大于约I OOPa,例如不小于约40Pa。 The composition value of G 'is typically between about 40Pa about IOOPa, such as not greater than about I OOPa, such as not less than about 40Pa.

[0021] 在本发明的另一方面,提供了治疗患者皮肤上的细纹的方法。 [0021] In another aspect of the present invention, there is provided a method of treating fine lines on the skin of a patient. 在一个实施方案中, 所述方法包括向患者的皮肤中引入包含透明质酸组分、交联所述透明质酸的交联组分和除所述交联组分以外的添加剂的混合物的组合物的步骤,所述组合物大体上光学透明,并且其中相对于除没有所述添加剂外,大体上相同的组合物,所述组合物表现出廷德尔效应减小。 In one embodiment, the method comprises introducing into a patient's skin comprising a hyaluronic acid component, a combination of a mixture of the crosslinked hyaluronic acid crosslinking components and additives other than the crosslinking component step thereof, the composition is substantially optically transparent, and wherein the additive does not except with respect to an outer, substantially the same composition, said composition exhibits reduced Tyndall effect.

[0022] 在本发明的另一方面,提供了改善面部美观的方法,所述方法通常包括下列步骤: 向患者的真皮区施用大体上光学透明的没有表现出或表现出不明显的廷德尔效应的真皮填充剂组合物。 [0022] In another aspect of the present invention, there is provided a method for improving surface appearance, the method generally comprises the steps of: administering to a dermal region of the patient a substantially optically transparent showed no Tyndall effect or exhibits insignificant dermal filler composition. 所述组合物通过下列步骤制成:提供透明质酸,使交联剂与维生素C衍生物反应,将已反应的交联剂与维生素C衍生物添加到所述透明质酸中以形成包括共价结合的维生素C的交联透明质酸组合物;并且匀化并中和所述交联透明质酸组合物以获得可注射的真皮填充剂组合物。 The composition is prepared by the steps of: providing hyaluronic acid, a crosslinking agent and vitamin C derivative, the crosslinking agent is added and reacted to the vitamin C derivative comprising hyaluronic acid to form a co- covalently bound vitamin C composition crosslinked hyaluronic acid; and homogenized and neutralizing the crosslinked hyaluronic acid composition to obtain an injectable dermal filler composition. 在一些实施方案中,维生素C衍生物是AA2G。 In some embodiments, the vitamin C derivative is AA2G. 在其它实施方案中,维生素C衍生物是维生食物。 In other embodiments, the vitamin C derivative is Vitagen.

[0023] 在本发明的又一方面,提供了减少患者薄皮区中细纹出现的方法,其中所述方法通常包括在不大于约Imm的深度,向所述患者施用真皮填充剂组合物,一种大体上光学透明的基于透明质酸的真皮填充剂组合物,其包括维生素C或维生素C衍生物。 [0023] In a further aspect of the present invention, there is provided a method of reducing the appearance of fine lines in the thin skin region of a patient, wherein said method generally comprising a depth of no more than about Imm, dermal filler composition is administered to the patient, a species substantially optically transparent hyaluronic acid-based dermal filler composition comprising vitamin C or a vitamin C derivative. 在一些实施方案中,在不大于〇. 8_、不大于0.6mm或不大于0.4mm的深度注射所述组合物。 In some embodiments, not greater than square. 8_, no greater than 0.6mm or no greater than the depth of injection of the composition of 0.4mm.

[0024] 在本发明的又一方面,提供了一种真皮填充剂组合物,其大体上光学透明并且通常包含与交联组分交联的透明质酸组分和与透明质酸组分共价结合的维生素C衍生物。 [0024] In a further aspect of the present invention, there is provided a dermal filler composition, which generally comprises a substantially optically transparent with hyaluronic acid component and a crosslinking component and a crosslinking component with a hyaluronic acid total vitamin C derivative covalently bonded. 在一个示例性实施方案中,所述组合物的G'值介于约40Pa和约IOOPa之间。 In one exemplary embodiment, the composition has a G 'value is between about 40Pa about IOOPa. 进一步地,所述组合物的透明质酸浓度可能介于约18mg/g和约30mg/g之间。 Further, the hyaluronic acid concentration of the composition may range between about 18mg / g and about 30mg / g. 在治疗皮肤,例如,甚至非常薄的皮肤,例如厚度不大于约Imm的皮肤上的细纹或浅表皱褶中,这些组合物可能特别有用且有效。 In the treatment of skin, for example, or even very thin skin, such as fine lines or thickness no greater than about Imm superficial wrinkles on the skin, these compositions may be particularly useful and effective. 在一些实施方案中,本发明的组合物在引入皮肤后持续至少3个月,至少6个月或长达一年。 In some embodiments, the compositions of the present invention after introduction into the skin for at least 3 months, or at least 6 months up to a year.

[0025] 参考下列附图和详细描述可能更容易理解和认识本发明的这些和其它方面及优点。 [0025] These and with reference to the following drawings and detailed description of other aspects and advantages may be more readily understood and appreciated the present invention.

[0026] 附图简述 [0026] BRIEF DESCRIPTION

[0027] 图1是L-抗坏血酸2-葡萄糖苷(AA2G)的结构示意图。 [0027] FIG. 1 is a schematic structural diagram of L- ascorbic acid 2-glucoside (of AA2G) a.

[0028] 图2是抗坏血酸3-氨丙基磷酸酯(维生食物)的结构示意图。 [0028] FIG. 2 is a schematic view of 3-aminopropyl phosphate (Vitagen) ascorbic acid.

[0029] 图3是抗坏血酸磷酸钠(AA2P)的结构示意图。 [0029] FIG. 3 is a schematic diagram sodium ascorbyl phosphate (AA2P) a.

[0030] 图4是1,4-丁二醇二缩水甘油醚(BDDE)的结构示意图。 [0030] FIG. 4 is a schematic view of 1,4-butanediol diglycidyl ether (BDDE) a.

[0031] 图5是季戊四醇缩水甘油醚(Star-PEG环氧化物)的结构示意图。 [0031] FIG. 5 is a schematic structural diagram of pentaerythritol ether (Star-PEG epoxide) of.

[0032] 图6是季戊四醇(3-氨丙基)醚(Star-PEG膨的结构示意图。 [0032] FIG. 6 is a pentaerythritol (3-aminopropyl) ether (Star-PEG structure diagram of swelling.

[0033] 图7是示出了根据本发明所述的各种真皮填充剂组合物的结合度和G'值的表。 [0033] FIG. 7 is a diagram illustrating a 'table of various values ​​in conjunction with the dermal filler composition of the present invention and G.

[0034] 图8是示出了根据本发明所述的HA-AA2G⑽DE)真皮填充剂组合物的结合度、HA浓度和G'值的表。 [0034] FIG. 8 is a table showing the HA-AA2G⑽DE according to the present invention) binding of the dermal filler composition, HA concentration and the G 'values.

[0035] 图9是对4种不同α-葡萄糖苷酶浓度而言,在时间方面(按分钟计),观察到的来自AA2G在PBS中的溶液的AsA释放的百分比的图形示意图。 [0035] FIG. 9 is different in terms of α- glucosidase four different concentrations, graphical schematic AsA release percentage in terms of time (in minutes), observed from a solution of AA2G in PBS.

[0036] 图10示出了来自根据本发明所述的结合真皮填充剂的游离AsA的释放曲线示意图(持续释放)(与反应时间相比的AA2G转化mo 1 %)。 [0036] FIG. 10 shows a schematic diagram of the release profile from the free AsA dermal filler binding according to the present invention (sustained release) (as compared with the reaction time of the conversion AA2G mo 1%).

[0037] 图IlA和IlB示出了根据本发明所述的各种真皮填充剂的附加释放数据。 [0037] FIG IlA and IlB release illustrate additional data according to various dermal filler according to the invention.

[0038] 图12示出了在浅表注射本发明基于HA的真皮填充凝胶和一些市场上可买到的供细纹应用的凝胶后,皮肤的图像。 [0038] FIG. 12 shows the gel after application of the supply lines of the present invention based on the injection of superficial dermal HA gel filled, and some commercially available, the skin image.

[0039] 图13示出了本发明基于HA的真皮填充凝胶和某些市场上可买到的供细纹应用的凝胶的视觉廷德尔得分。 [0039] FIG. 13 shows a gel of the present invention based on the supply lines HA dermal application of gel filled and certain commercially available Tyndall visual score.

[0040] 图14示出了本发明基于HA的真皮填充凝胶和一些市场上可买到的供细纹应用的凝胶从皮肤出射的蓝光%。 [0040] FIG. 14 shows a gel of the present invention is based on the supply lines HA dermal application, and some gel-filled commercially available blue light emitted from the skin%.

[0041] 图15示出了植入本发明基于HA的真皮填充凝胶和一些市场上可买到的供细纹应用的凝胶1周后,剩余凝胶的总%。 [0041] FIG. 15 illustrates the implantation of the gel for the present invention is based on the application of fine lines on the gel HA dermal filler, and some commercially available one week, the total% of the remaining gel.

[0042] 图16示出了在植入本发明基于HA的真皮填充凝胶和一些市场上可买到的供细纹应用的凝胶第0周、第12周、第24周和第40周,剩余凝胶的总%。 [0042] FIG. 16 shows the gel of the present invention implanted for the application lines HA-based dermal filler gels and the number of commercially available Week 0 Week 12 Week 24 and Week 40 the total% of the remaining gel.

[0043] 详述 [0043] detail

[0044] 在本发明的一个方面,提供了真皮填充剂组合物,所述组合物通常包含生物相容性聚合物,例如多糖(例如交联透明质酸)和与所述聚合物共价结合的维生素C衍生物。 [0044] In one aspect of the invention there is provided a dermal filler composition, the composition typically comprises a biocompatible polymer, such as polysaccharides (e.g. cross-linked hyaluronic acid) and covalently bound to the polymer vitamin C derivative. 所述组合物提供了持续释放供皮肤胶原蛋白增生的维生素C以及其它治疗或美容利益。 The composition provides sustained release of the skin collagen for vitamin C, and other therapeutic or cosmetic benefit. 当引入皮肤时,例如皮内,所述组合物与体内的内源性酶反应,并且随时间推移,经酶裂解在体内生成生物活性维生素C。 When introduced into the skin, e.g. intradermal, said reactant composition with endogenous enzymes in the body, and over time, generated by enzymatic cleavage in vivo biological activity of vitamin C. 因为在几周或几个月内维生素C从所述组合物释放,使得随之而来的利益对身体可用。 Since the release from the composition in a few weeks or months of vitamin C, so that the attendant benefits available to the body.

[0045] 所述聚合物可选自蛋白质、肽和多肽、聚赖氨酸、胶原蛋白、前胶原蛋白、弹性蛋白和层粘连蛋白。 The [0045] polymer may be selected from proteins, peptides and polypeptides, polylysine, collagen, procollagen, laminin and elastin.

[0046] 所述聚合物可选自具有羟基、胺和羧基官能团的合成聚合物:聚(乙烯醇)、聚乙二醇、聚乙烯胺、聚烯丙胺、去乙酰基聚丙烯酰胺、聚丙烯酸和聚甲基丙烯酸。 [0046] The polymer may be selected from synthetic polymers having hydroxyl, amine and carboxyl functional groups: poly (vinyl alcohol), polyethylene glycol, polyvinyl amine, polyallylamine, deacetyl polyacrylamide, polyacrylic acid and polymethacrylic acid. 所述聚合物可选自树枝状或支化聚合物,包括树枝状多元醇和树枝状聚胺。 The polymer may be selected from a dendritic or branched polymers, dendrimers comprising dendritic polyols and polyamines. 所述聚合物可选自具有羟基、胺和羧基官能团的固体表面。 The polymer may be selected from a solid surface having hydroxyl, amine and carboxyl functional groups.

[0047] 所述聚合物可为多糖,例如选自下列组的多糖,包括淀粉及其衍生物;葡聚糖及其衍生物;纤维素及其衍生物;几丁质、壳聚糖和藻酸盐及其衍生物。 [0047] The polymer may be a polysaccharide, for example selected from the group of polysaccharides, including starch and derivatives thereof; dextran and derivatives thereof; cellulose and its derivatives; chitin, chitosan and alginic salts and derivatives thereof.

[0048] 在本发明的一个示例性实施方案中,所述聚合物为糖胺聚糖。 [0048] In one exemplary embodiment of the present invention, the polymer is a glycosaminoglycan. 本文公开的水凝胶组合物可进一步包含两种或更多种不同的糖胺聚糖聚合物。 Herein disclosed hydrogel composition may further comprise two or more different glycosaminoglycan polymer. 如本文所使用,术语“糖胺聚糖”与“GAG”和“粘多糖”同义并且指重复二糖单元组成的长直链多糖。 As used herein, the term "glycosaminoglycan" and "GAG" and "glycosaminoglycans" are used synonymously and refers to a repeating disaccharide units composed of long linear polysaccharide. 重复单元由与己糖胺(含氮的六碳糖)及其药学上可接受的盐连接的己糖(六碳糖)或己糖醛酸组成。 And hexosamine repeating unit represented by (hexose nitrogen) and pharmaceutically acceptable hexoses (six carbon sugar) or salts linked hexuronic acid. GAG家族成员在其所含的己糖胺、己糖或己糖醛酸单元的类型上不同,例如葡萄糖醛酸、艾杜糖醛酸、 半乳糖、半乳糖胺、葡萄糖胺并且在配糖键的几何形态上也可能不同。 GAG family contained on the type of hexosamine, hexose or hexuronic acid units of different, e.g. glucuronic acid, iduronic acid, galactose, galactosamine, glucosamine and the glycosidic bond it may be different on geometry. 任何糖胺聚糖聚合物在本文公开的水凝胶组合物中均有用,条件是糖胺聚糖聚合物改善皮肤状况。 Any glycosaminoglycan polymer used herein are disclosed in the hydrogel composition, with the proviso that glycosaminoglycan polymer improving skin condition. 糖胺聚糖的非限制性实例包括硫酸软骨素、硫酸皮肤素、硫酸角质素、透明质烷。 Non-limiting examples of glycosaminoglycans include chondroitin sulfate, dermatan sulfate, keratan sulfate, hyaluronan. 糖胺聚糖可接受的盐的非限制性实例包括钠盐、钾盐、镁盐、钙盐及其组合。 Glycosaminoglycans Nonlimiting examples of acceptable salts include sodium, potassium, magnesium, calcium, and combinations thereof. 例如,在Piron和Thol in, Polysaccharide Crosslinking?HydrogeI Preparation,Resulting Polysaccharides (s) and Hydrogel(s),uses Thereof,美国专利公布2003/0148995;Lebreton,Cross-Linking of Low and High Molecular Weight Polysaccharides Preparation of Injectable Monophase Hydrogels;Lebreton,Viscoelastic Solutions Containing Sodium Hyaluronate and Hydroxypropyl Methyl Cellulose?Preparation and Uses,美国专利公布2008/0089918;Lebreton,Hyaluronic Acid-Based Gels Including Lidocaine,美国专利公布2010/0028438;和Polysaccharides and Hydrogels thus Obtained,美国专利公布2006/0194758;和Di Napoli,Composition and Method for Intradermal Soft Tissue Augmentation,国际专利公布WO 2004/073759中描述了在本文公开的水凝胶组合物和方法中有用的糖胺聚糖及其所得聚合物,其各自据此通过引用整体并入。 ? For example, in Piron and Thol in, Polysaccharide Crosslinking HydrogeI Preparation, Resulting Polysaccharides (s) and Hydrogel (s), uses Thereof, US Patent Publication 2003/0148995; Lebreton, Cross-Linking of Low and High Molecular Weight Polysaccharides Preparation of Injectable Monophase Hydrogels;? Lebreton, Viscoelastic Solutions Containing Sodium Hyaluronate and Hydroxypropyl Methyl Cellulose Preparation and Uses, US Patent publication 2008/0089918; Lebreton, Hyaluronic Acid-Based Gels Including Lidocaine, US Patent publication 2010/0028438; and Polysaccharides and Hydrogels thus Obtained, U.S. Patent Publication 2006/0194758; and Di Napoli, composition and method for Intradermal Soft Tissue Augmentation, international Patent Publication WO 2004/073759 describes hydrogel compositions useful in the compositions and methods herein disclosed and glycosaminoglycans The resulting polymer, each of which is hereby incorporated by reference. 在本文公开的水凝胶组合物和方法中有用的GAG在市场上可买到,例如基于透明质烷的真皮填充剂JUVEDERVr > JUVEDERMX30, JUVEDERM'; U11 ra , JUVEDERNf Ultra Plus、JUVEDERM⑧Ultra XC和JUVEDERM⑨Ultra Plus XC (Allergan Inc,Irvine, California)。 In the hydrogel composition and methods disclosed herein are useful in GAG commercially available, for example, fillers JUVEDERVr> JUVEDERMX30, JUVEDERM 'based on hyaluronan leather; U11 ra, JUVEDERNf Ultra Plus, JUVEDERM⑧Ultra XC and JUVEDERM⑨Ultra Plus XC (Allergan Inc, Irvine, California). 表1列出了代表性GAG。 Table 1 lists representative GAG.

Figure CN108379112AD00081

[0050]本发明的方面部分地提供了包含硫酸软骨素聚合物的水凝胶组合物。 [0050] aspect of the present invention provide, in part hydrogel composition comprising a polymer of chondroitin sulfate. 如本文所使用,术语“硫酸软骨素聚合物”指可变长度的无支链的、硫酸化聚合物,其包含两个交替单糖即D-葡萄糖醛酸(GlcA)和N-乙酰基-D-半乳糖胺(GalNAc)及其药学上可接受的盐的二糖。 As used herein, the term "chondroitin sulfate polymer" refers to, sulfated polymers unbranched of variable length, comprising two alternating monosaccharides i.e. D- glucuronic acid (GIcA) and N- acetyl - D- galactosamine (GalNAc) and disaccharide pharmaceutically acceptable salt thereof. 硫酸软骨素聚合物也可包括差向异构化为L-艾杜糖醛酸(IdoA)的D-葡萄糖醛酸残基,在这种情况下将所得二糖称为硫酸皮肤素。 Chondroitin sulfate polymers may also include L- epimerized to iduronic acid (IdoA) of D- glucuronic acid residue, in this case referred to as the resultant disaccharide dermatan sulfate. 硫酸软骨素可具有超过100个单独糖的链,其各自以可变位置和量硫酸化。 Chondroitin sulfate can have over 100 individual sugars chains, each variable position and of the amount of sulfuric acid. 硫酸软骨素聚合物是软骨的重要结构组分并且提供了其对压缩的许多抗性。 Chondroitin sulfate is an important structural component of cartilage polymer and provides many of its resistance to compression. 任何硫酸软骨素聚合物在本文公开的组合物中均有用,条件是硫酸软骨素聚合物改善皮肤状况。 Any polymer are chondroitin sulfate used in the compositions disclosed herein, with the proviso that the polymer chondroitin sulfate improving skin condition. 硫酸软骨素药学上可接受的盐的非限制性实例包括硫酸软骨素钠、硫酸软骨素钾、硫酸软骨素镁、硫酸软骨素轉及其组合。 Chondroitin sulfate pharmaceutically acceptable, non-limiting examples of salts include sodium chondroitin sulfate, potassium chondroitin sulfate, magnesium sulfate, chondroitin, chondroitin sulfate, and combinations thereof turn.

[0051] 本说明书的方面部分地提供了包含硫酸角质素聚合物的水凝胶组合物。 Aspect of the [0051] section of the present specification provides a hydrogel composition comprising a polymer keratan sulfate. 如本文所使用,术语“硫酸角质素聚合物”指包含二糖单元的可变长度的聚合物,其本身包括β-D-半乳糖和N-乙酰基-D-半乳糖胺(GalNAc)及其药学上可接受的盐。 As used herein, the term "keratan sulfate polymer" refers to polymers of variable length comprising disaccharide units, which itself comprises β-D- galactose and N- acetyl -D- galactosamine (GalNAc) and a pharmaceutically acceptable salt thereof. 硫酸角质素重复区内的二糖可能经岩藻糖基化并且N-乙酰神经氨酸盖在链的末端。 Keratan sulfate disaccharide repeat region may be glycosylated fucose and by N- terminal cover acetylneuraminic acid chains. 任何硫酸角质素聚合物在本文公开的组合物中均有用,条件是硫酸角质素聚合物改善皮肤状况。 Any keratan sulfate used herein disclosed are polymer compositions, with the proviso that the polymer keratan sulfate improving skin condition. 硫酸角质素药学上可接受的盐的非限制性实例包括硫酸角质素钠、硫酸角质素钾、硫酸角质素镁、硫酸角质素钙及其组合。 Keratan sulfate pharmaceutically acceptable non-limiting examples of salts include sodium prime keratin sulfate, keratan sulfate, potassium, magnesium keratan sulfate, keratan sulfate, calcium, and combinations thereof.

[0052] 本说明书的方面部分地提供了包含透明质烷聚合物的水凝胶组合物。 Aspect of the [0052] section of the present specification provides a hydrogel composition comprising hyaluronan polymer. 如本文所使用,术语“透明质烷聚合物”与“HA聚合物”、“透明质酸聚合物”和“透明质酸盐聚合物”同义, 指包含二糖单元的阴离子、非硫酸化糖胺聚糖聚合物,其本身包括通过交替的β_1,4和β-1, 3糖苷键连接在一起的D-葡萄糖醛酸和DN-乙酰葡萄糖胺单体及其药学上可接受的盐。 As used herein, the term "hyaluronan polymer" and "HA polymer", "hyaluronic acid polymer" and "polymeric hyaluronate" are synonymous, refers disaccharide units containing an anionic, non-sulfated glycosaminoglycan polymer, which itself comprises a pharmaceutically linked together by alternating β_1,4 and β-1, 3 glycosidic bonds D- glucuronic acid and DN- acetylglucosamine monomers and pharmaceutically acceptable salts thereof. 可从动物和非动物来源纯化透明质烷聚合物。 It can be purified from animal and non-animal origin hyaluronan polymer. 透明质烷的聚合物尺寸范围可从约5,000Da到约20,000,000Da。 Polymeric hyaluronan can range in size from about 5,000Da to about 20,000,000Da. 任何透明质烷聚合物在本文公开的组合物中均有用,条件是透明质烷改善皮肤状况。 Any hyaluronan polymer disclosed herein are used in the compositions, with the proviso that hyaluronan improving skin condition. 透明质烷药学上可接受的盐的非限制性实例包括透明质烷钠、透明质烷钾、透明质烷镁、透明质烷钙及其组合。 Hyaluronan pharmaceutically acceptable non-limiting examples of salts include sodium hyaluronan, hyaluronan, potassium, magnesium hyaluronan, hyaluronan calcium, and combinations thereof.

[0053] 本说明书的方面部分地提供了包含交联糖胺聚糖聚合物的水凝胶组合物。 [0053] aspect of the specification provides a part hydrogel composition comprising a crosslinked glycosaminoglycan polymer. 如本文所使用,术语“交联”指分子间键将单独的聚合物分子或单体链连接成像凝胶一样更稳定的结构。 As used herein, the term "crosslinked" refers to intermolecular bonds the individual polymer molecules or monomers as the chain connector forming a more stable gel structure. 因而,交联糖胺聚糖聚合物具有将至少一个单独聚合物分子连接到另一个的至少一个分子间键。 Thus, crosslinked glycosaminoglycan polymer having at least one intermolecular bond at least a single polymer molecule to another. 糖胺聚糖聚合物的交联通常导致水凝胶形成。 Crosslinked glycosaminoglycan polymer typically results in formation of a hydrogel. 此类水凝胶粘度高并且需要相当大的力挤出细针。 Such hydrogel adhesive is high and requires considerable force extrusion needle. 可使用二醛和二硫化物交联剂交联本文公开的糖胺聚糖聚合物,包括但不限于多功能PEG基交联剂、二乙烯砜、缩水甘油醚和双环氧化物、双碳二亚胺。 Using dialdehyde disulfide glycosaminoglycan polymer and crosslinking agent disclosed herein, including but not limited to PEG-based multifunctional crosslinking agent, divinyl sulfone, ether, and diepoxide, bis carbodiimide imide. 透明质烷交联剂的非限制性实例包括多功能PEG基交联剂,如季戊四醇四缩水甘油醚(PETGE)、二乙烯砜(DVS)、1,4-丁二醇二缩水甘油醚(BDDE)、1,2-双(2,3-环氧丙氧基)乙烯(E⑶GE)、1,2, 7,8—二环氧辛烷(DEO)、(亚苯基双-(乙基)-碳二亚胺和1,6亚己基双(乙基碳二亚胺)、己二酰肼(ADH)、双(磺基琥珀基)辛二酸酯(BS)、己二胺(HMDA)、1_ (2,3-环氧丙氧基)-2,3-环氧环己烧、赖氨酸、赖氨酸甲酯或其组合。在Stroumpoulis和Tezel,Tunably Crosslinked Polysaccharide Compositions,2010年10月22日提交的美国专利申请12/910,466中公开了其它有用的交联剂,其通过引用整体并入。例如,在Piron和ThoIin,Polysaccharide Crosslinking,Hydrogel Preparation,Resulting Polysaccharides (s)and Hydrogel (s),uses Thereof,美国专利公布2003/0148995;Lebreton,Cross-Linking of Low and High Molecular Weight Polysaccharides Preparation of Injectable Monophase Non-limiting examples of crosslinking agents include hyaluronan PEG multifunctional cross-linking agent, such as pentaerythritol tetraglycidyl ether (of PETGE), divinyl sulfone (DVS), 1,4- butanediol diglycidyl ether (BDDE ), 1,2-bis (2,3-epoxypropoxy) ethylene (E⑶GE), 1,2, 7,8- diepoxyoctane (DEO), (phenylene bis - (ethyl) - carbodiimide and 1,6-hexamethylene bis (ethylcarbodiimide), adipic dihydrazide (of ADH), bis (sulfosuccinimidyl-yl) suberate (the BS), hexamethylene diamine (of HMDA) , 1_ (2,3-epoxypropoxy) -2,3-epoxycyclohexyl burning, lysine, lysine methyl ester, or combinations thereof. in Stroumpoulis and Tezel, Tunably Crosslinked Polysaccharide compositions, 2010 October U.S. Patent application filed May 22, 12 / 910,466 discloses other useful crosslinking agents, which is incorporated by reference in its entirety. For example, in Piron and ThoIin, Polysaccharide crosslinking, Hydrogel Preparation, Resulting Polysaccharides (s) and Hydrogel (s), uses Thereof, U.S. Patent publication 2003/0148995; Lebreton, Cross-Linking of Low and High Molecular Weight Polysaccharides Preparation of Injectable Monophase Hydrogels;Lebreton,Viscoelastic Solutions Containing Sodium Hyaluronate and Hydroxypropyl Methyl Cellulose,Preparation and Uses,美国专利公布2008/0089918; Lebreton,Hyaluronic Acid-Based Gels Including Lidocaine,美国专利公布2010/ 0028438;和Polysaccharides and Hydrogels thus Obtained,美国专利公布2006/ 0194758;和Di Napoli,Composition and Method for Intradermal Soft Tissue Augmentation,国际专利公布WO 2004/073759中描述了交联糖胺聚糖聚合物的方法的非限制性实例,以及在本文公开的组合物和方法中有用的糖胺聚糖聚合物,其各自据此通过引用整体并入。 Hydrogels; Lebreton, Viscoelastic Solutions Containing Sodium Hyaluronate and Hydroxypropyl Methyl Cellulose, Preparation and Uses, US Patent Publication 2008/0089918; Lebreton, Hyaluronic Acid-Based Gels Including Lidocaine, US Patent Publication 2010/0028438; and Polysaccharides and Hydrogels thus Obtained, United States Patent Publication 2006/0194758; and Di Napoli, Composition and method for Intradermal Soft Tissue Augmentation, international Patent Publication describes a non-limiting example of a method of the crosslinked glycosaminoglycan polymer 2004/073759 WO, and disclosed herein the compositions and methods useful glycosaminoglycan polymer, each of which is hereby incorporated by reference.

[0054] 本说明书的方面部分地提供了包含具有一定交联度的交联糖胺聚糖聚合物的水凝胶组合物。 Aspect of the [0054] section of the present specification provides a hydrogel composition comprising a crosslinked glycosaminoglycan polymer having a certain degree of crosslinking. 如本文所使用,术语“交联度”指糖胺聚糖聚合物单体单元,例如与交联剂结合的透明质烷二糖单体单元的百分比。 As used herein, the term "degree of crosslinking" refers to a glycosaminoglycan polymer monomeric units, for example, the percentage of hyaluronan disaccharide monomer units in combination with the crosslinking agent. 交联度表示为交联剂与糖胺聚糖的重量百分比。 The degree of crosslinking expressed as a weight percentage of the crosslinking agent glycosaminoglycan. 在本发明的某些有利实施方案中,交联度介于约3%和约12%之间,例如介于约5 %和约10%之间。 In certain advantageous embodiments of the invention, the degree of crosslinking of between about 3% and about 12%, for example between about 5% and about 10%.

[0055] 在一个实施方案中,水凝胶组合物包含交联糖胺聚糖聚合物,例如交联透明质酸, 其中所述组合物中存在的所述交联糖胺聚糖聚合物的浓度(例如)介于约18mg/g和约30mg/ g之间。 [0055] In one embodiment, the hydrogel composition comprising a crosslinked glycosaminoglycan polymer, such as cross-linked hyaluronic acid, wherein the composition is present in the crosslinked glycosaminoglycan polymer concentration (e.g.) between about 18mg / g and about 30mg / g. 在一些实施方案中,所述组合物的透明质酸总浓度为约24mg/g或约25mg/g。 In some embodiments, the total concentration of the hyaluronic acid of the composition is from about 24mg / g or from about 25mg / g.

[0056] 本说明书的方面部分地提供了包含低分子量的透明质烷聚合物、高分子量的透明质烷聚合物或低分子量和高分子量的透明质烷聚合物的水凝胶组合物。 Aspect [0056] This section provides a description of hyaluronan comprising a low molecular weight polymer, a high molecular weight hydrogel composition hyaluronan or a hyaluronan polymer of a low molecular weight polymer and a high molecular weight. 如本文所使用,当提及“透明质烷”时术语“高分子量”指平均分子量为l,〇〇〇,〇〇〇Da或更高的透明质烷聚合物。 As used herein, when referring to "hyaluronan" The term "high molecular weight" refers to the average molecular weight of L, 〇〇〇, 〇〇〇Da or higher hyaluronan polymer. 高分子量透明质烷聚合物的非限制性实例包括约1,500,OOODa、约2,000,OOODa、约2, 500,OOODa、约3,000,OOODa、约3,500,OOODa、约4,000,OOODa、约4,500,OOODa和约5,000, OOODa的透明质烷聚合物。 High molecular weight hyaluronan polymer Non-limiting examples include about 1,500, OOODa, about 2,000, OOODa, about 2, 500, OOODa, about 3,000, OOODa, about 3,500, OOODa, about 4 , 000, OOODa, about 4,500, OOODa about 5,000, OOODa hyaluronan polymer. 如本文所使用,当提及“透明质烧”时术语“低分子量”指平均分子量小于l,000,000Da的透明质烷聚合物。 As used herein, when referring to "the hyaluronic burn" The term "low molecular weight" refers to the average molecular weight is less than L, the hyaluronan polymer 000,000Da. 低分子量透明质烷聚合物的非限制性实例包括约200,00003、约300,00003、约400,00003、约500,00003、约600,00003、约700,00003、约800, OOODa和约900,OOODa的透明质烷聚合物。 Non-limiting examples of hyaluronan polymer comprising a low molecular weight of about 200,00003, about 300,00003, about 400,00003, about 500,00003, about 600,00003, about 700,00003, about 800, OOODa about 900, OOODa hyaluronan polymer.

[0057] 在一个实施方案中,组合物包含低分子量的交联透明质烷聚合物。 [0057] In one embodiment, the composition comprises a crosslinked hyaluronan low molecular weight polymers. 在该实施方案的方面,组合物包含平均分子量为(例如)约100,OOODa、约200,OOODa、约300,OOODa、约400, OOODa、约500,0000&、约600,0000&、约700,0000&、约800,0000&或约900,0000&的交联透明质烷聚合物。 In aspects of this embodiment, the composition comprises an average molecular weight of (e.g.) about 100, OOODa, about 200, OOODa, about 300, OOODa, about 400, OOODa, about 500,0000 & amp ;, from about 700 to about 600,0000 & amp ;, , 0000 & amp ;, about 800,0000 & amp; or from about 900,0000 & amp; crosslinked hyaluronan polymer. 在该实施方案的其它方面,组合物包含平均分子量为(例如)至多l〇〇,〇〇〇Da、 至多200,00003、至多300,00003、至多400,00003、至多500,00003、至多600,00003、至多700,OOODa、至多800,OOODa、至多900,OOODa或至多950,OOODa的交联透明质烷聚合物。 In other aspects of this embodiment, the composition comprises an average molecular weight of (e.g.) up l〇〇, 〇〇〇Da, most 200,00003, most 300,00003, most 400,00003, most 500,00003, at most 600, 00003, up to 700, OOODa, up to 800, OOODa, up to 900, OOODa or up to 950, OOODa crosslinked hyaluronan polymer. 在该实施方案的其它方面,组合物包含平均分子量为(例如)约100,OOODa至约500,OOODa、约200,OOODa 至约500,000Da、约300,OOODa 至约500,000Da、约400,OOODa 至约500,000Da、约500,OOODa 至约950,000Da、约600,OOODa 至约950,000Da、约700,OOODa 至约950,000Da、约800,OOODa 至约950,000Da、约300,OOODa 至约600,000Da、约300,OOODa 至约700,000Da、约300,OOODa至约800,OOODa或约400,OOODa至约700,OOODa的交联透明质烷聚合物。 In other aspects of this embodiment, the composition comprises an average molecular weight of (e.g.) about 100, OOODa to about 500, OOODa, about 200, OOODa to about 500,000 Da, about 300, OOODa to about 500,000 Da, about 400, OOODa to about 500,000 Da, about 500, OOODa to about 950,000 Da, about 600, OOODa to about 950,000 Da, about 700, OOODa to about 950,000 Da, about 800, OOODa to about 950,000 Da, about 300, OOODa to about 600,000Da, about 300, OOODa to about 700,000Da, about 300, OOODa to about 800, OOODa or about 400, OOODa to about 700, OOODa crosslinked hyaluronan polymer.

[0058] 在另一实施方案中,组合物包含高分子量的交联透明质烷聚合物。 [0058] In another embodiment, the composition comprises a high molecular weight crosslinked hyaluronan polymer. 在该实施方案的方面,组合物包含平均分子量为(例如)约1,〇〇〇,OOODa、约1,500,OOODa、约2,000,OOODa、 约2,500,OOODa、约3,000,OOODa、约3,500,OOODa、约4,000,OOODa、约4,500,OOODa或约5, 000,OOODa的交联透明质烷聚合物。 In aspects of this embodiment, the composition comprises an average molecular weight of (e.g.) about 1, 〇〇〇, OOODa, about 1,500, OOODa, about 2,000, OOODa, about 2,500, OOODa, about 3,000 , OOODa, about 3,500, OOODa, about 4,000, OOODa, about 4,500, OOODa or about 5, 000, OOODa crosslinked hyaluronan polymer. 在该实施方案的其它方面,组合物包含平均分子量为(例如)至少1,000,00003、至少1,500,00003、至少2,000,00003、至少2,500,00003、至少3, 000,000Da、至少3,500,000Da、至少4,000,000Da、至少4,500,OOODa 或至少5,000,OOODa 的交联透明质烷聚合物。 In other aspects of this embodiment, the composition comprises an average molecular weight of (e.g.) at least 1,000,00003 least 1,500,00003 least 2,000,00003 least 2,500,00003, at least 3, 000,000Da, at least 3 , 500,000Da, at least 4,000,000Da, at least 4,500, OOODa or at least 5,000, OOODa crosslinked hyaluronan polymer. 在该实施方案的其它方面,组合物包含平均分子量为(例如)约1, 000,OOODa 至约5,000,000Da、约1,500,OOODa 至约5,000,000Da、约2,000,OOODa 至约5,000, 000Da、约2,500,OOODa 至约5,000,000Da、约2,000,OOODa 至约3,000,000Da、约2,500,OOODa 至约3,000,OOODa的交联透明质烷聚合物。 In other aspects of this embodiment, the composition comprises an average molecular weight of (e.g.) about 1, 000, OOODa to about 5,000,000 Da, about 1,500, OOODa to about 5,000,000 Da, about 2,000, OOODa to about 5,000, 000Da, about 2,500, OOODa to about 5,000,000Da, about 2,000, OOODa to about 3,000,000Da, about 2,500, OOODa to about 3,000, OOODa crosslinked hyaluronan polymer.

[0059] 又一实施方案中,组合物包含交联透明质烷聚合物,其中所述交联透明质烷聚合物包含不同比例的高分子量透明质烷聚合物和低分子量透明质烷聚合物的组合。 [0059] In yet another embodiment, the composition comprises a crosslinked hyaluronan polymer, wherein said crosslinked high molecular weight hyaluronan polymers containing different proportions of hyaluronan polymer and a low molecular weight polymer is hyaluronan combination. 在该实施方案的方面,组合物包含交联透明质烷聚合物,其中所述交联透明质烷聚合物包含比例为约20:1、约15:1、约10:1、约5:1、约1:1、约1:5、约1:10、约1:15或约1:20的高分子量透明质烷聚合物和低分子量透明质烷聚合物的组合。 In aspects of this embodiment, the composition comprises a crosslinked hyaluronan polymer, wherein the crosslinked hyaluronan polymer comprises a ratio of about 20: 1, about 15: 1, about 10: 1, about 5: 1 , from about 1: 1, about 1: 5, about 1:10, about 1:15, or high molecular weight hyaluronan about 1:20 and low-molecular weight polymer composition alkoxy hyaluronan polymer.

[0060] 本说明书的方面部分地提供了包含未交联糖胺聚糖聚合物的水凝胶组合物。 Aspect of the [0060] section of the present specification provides a hydrogel composition comprising a glycosaminoglycan linked polymer is uncrosslinked. 如本文所使用,术语“未交联”指缺乏连接单独的糖胺聚糖聚合物分子或单体链的分子间键。 As used herein, the term "non-crosslinked" refers to a lack of connectivity between individual monomer molecules or glycosaminoglycan polymer chain molecular bonds. 因而,未交联糖胺聚糖聚合物未通过分子间键与任何其它糖胺聚糖聚合物连接。 Thus, an uncrosslinked glycosaminoglycan polymer does not bond to any other glycosaminoglycan polymer by intermolecular. 在该实施方案的方面,组合物包含未交联硫酸软骨素聚合物、未交联硫酸皮肤素聚合物、未交联硫酸角质素聚合物、未交联乙酰肝素聚合物、未交联硫酸乙酰肝素聚合物或未交联透明质烷聚合物。 In aspects of this embodiment, the composition comprises non-cross-linked chondroitin sulfate polymer, uncrosslinked polymer skin-linked sulfate, keratin uncross-linked polymer sulfate, heparan uncrosslinked polymer, uncross-linked acetyl sulfate heparin or uncrosslinked polymer hyaluronan polymer. 未交联糖胺聚糖聚合物可溶于水并且通常在本质上保持流动性。 Uncrosslinked glycosaminoglycan polymer soluble in water and generally remain fluid in nature. 因而,未交联糖胺聚糖聚合物常与作为润滑剂的基于糖胺聚糖聚合物的水凝胶组合物混合以促进组合物通过细针的挤出过程。 Thus, an uncrosslinked glycosaminoglycan polymer as a lubricant is often mixed with a hydrogel composition based on glycosaminoglycan polymer extrusion process to facilitate the composition through a fine needle.

[0061] 在一个实施方案中,组合物包含未交联糖胺聚糖聚合物,其中所述未交联糖胺聚糖聚合物存在的浓度为(例如)约2mg/g、约3mg/g、约4mg/g、约5mg/g、约6mg/g、约7mg/g、约8mg/g、约9mg/g、约IOmg/g、约Ilmg/g、约12mg/g、约13mg/g、约13·5mg/g、约14mg/g、约15mg/ g、约16mg/g、约17mg/g、约18mg/g、约19mg/g、约20mg/g、约40mg/g 或约60mg/g。 [0061] In one embodiment, the composition comprises cross-linked non-glycosaminoglycan polymer, wherein said uncrosslinked polymer is present in a concentration of glycosaminoglycan linked to (e.g.) about 2mg / g, from about 3mg / g , about 4mg / g, from about 5mg / g, from about 6mg / g, from about 7mg / g, from about 8mg / g, from about 9mg / g, from about IOmg / g, from about Ilmg / g, from about 12mg / g, from about 13mg / g , from about 13 · 5mg / g, from about 14mg / g, from about 15mg / g, from about 16mg / g, from about 17mg / g, from about 18mg / g, from about 19mg / g, from about 20mg / g, from about 40mg / g or from about 60mg / g. 在该实施方案的其它方面,组合物包含未交联糖胺聚糖,其中所述未交联糖胺聚糖存在的浓度为(例如)至少lmg/g、至少2mg/g、至少3mg/g、至少4mg/g、至少5mg/g、至少I Omg/g、至少15mg/g、至少20mg/g、至少25mg/g、至少35mg/g或至少40mg/g。 In other aspects of this embodiment, the composition comprises an uncrosslinked glycosaminoglycan, wherein said non-cross-linked glycosaminoglycan present in a concentration of (e.g.) at least lmg / g, at least 2mg / g, at least 3mg / g at least 4mg / g, at least 5mg / g, at least I Omg / g, at least 15mg / g, at least 20mg / g, at least 25mg / g, at least 35mg / g, or at least 40mg / g. 在该实施方案的其它方面,组合物包含未交联糖胺聚糖,其中所述未交联糖胺聚糖存在的浓度为(例如)至多lmg/g、至多2mg/g、至多3mg/g、至多4mg/g、至多5mg/g、至多I Omg/g、至多15mg/g、至多20mg/g或至多25mg/g。 In other aspects of this embodiment, the composition comprises an uncrosslinked glycosaminoglycan, wherein said non-cross-linked glycosaminoglycan present in a concentration of (e.g.) up to lmg / g, up to 2mg / g, up to 3mg / g , up to 4mg / g, up to 5mg / g, at most I Omg / g, up to 15mg / g, up to 20mg / g or up to 25mg / g. 在该实施方案的其它方面,组合物包含未交联糖胺聚糖,其中所述未交联糖胺聚糖存在的浓度为(例如)约lmg/g至约60mg/g、约I Omg/g至约40mg/g、约7 · 5mg/g至约19 · 5mg/g、约8 · 5mg/g 至约18 · 5mg/g、约9 · 5mg/g至约17 · 5mg/g、约10 · 5mg/g至约16 · 5mg/g、约11 · 5mg/g至约15 · 5mg/g或约12 · 5mg/g至约14 · 5mg/g。 In other aspects of this embodiment, the composition comprises an uncrosslinked glycosaminoglycan, wherein the presence of the uncrosslinked glycosaminoglycan concentration (e.g.) about lmg / g to about 60mg / g, about I Omg / g to about 40mg / g, about 7 · 5mg / g to about 19 · 5mg / g, about 8 · 5mg / g to about 18 · 5mg / g, about 9 · 5mg / g to about 17 · 5mg / g, about 10 · 5mg / g to about 16 · 5mg / g, from about 11 · 5mg / g to about 15 · 5mg / g or from about 12 · 5mg / g to about 14 · 5mg / g.

[0062] 本说明书的方面部分地提供了基本上不含交联糖胺聚糖聚合物的水凝胶组合物。 Aspect of the [0062] section of the present specification provides a substantially free of a crosslinked glycosaminoglycan polymer hydrogel composition. 如本文所使用,术语“基本上不含”(或“基本上由......组成”)指其中仅可检测到微量的交联母体聚合物的组合物。 As used herein, the term "substantially free" (or "consisting essentially of ......"), which only refers to a composition detectable traces of crosslinked polymer matrix. 在该实施方案的一个方面,组合物包含基本上不含交联硫酸软骨素聚合物的硫酸软骨素、基本上不含交联硫酸皮肤素聚合物的硫酸皮肤素、基本上不含交联硫酸角质素聚合物的硫酸角质素、基本上不含交联乙酰肝素聚合物的乙酰肝素、基本上不含交联硫酸乙酰肝素聚合物的硫酸乙酰肝素或基本上不含交联透明质烷聚合物的硫酸透明质烷。 In one aspect of this embodiment, the composition comprising substantially free of cross-linked chondroitin sulfate polymer, chondroitin sulfate, dermatan sulfate is substantially free of a crosslinked polymer with dermatan sulfate, sulfuric acid is substantially free of cross-linking keratan polymer keratan sulfate, heparan substantially free of cross-linked polymer heparan, heparan sulfate substantially free of a crosslinked polymer with heparan sulfate or substantially free of a crosslinked hyaluronan polymer sulfated hyaluronan.

[0063] 本说明书的方面部分地提供了完全不含交联糖胺聚糖聚合物的水凝胶组合物。 [0063] Aspects of the present specification provides a fully free of a crosslinked glycosaminoglycan polymer hydrogel composition. 如本文所使用,术语“完全不含”指组合物在使用的仪器或工艺检测范围内,不能检测到交联糖胺聚糖聚合物或不能确认其存在。 As used herein, the term "completely free" means that the composition or process within the detection range of the instrument used, can not be detected crosslinked glycosaminoglycan polymer or its presence can not be confirmed. 在该实施方案的一个方面,组合物包含完全不含交联硫酸软骨素聚合物的硫酸软骨素、完全不含交联硫酸皮肤素聚合物的硫酸皮肤素、完全不含交联硫酸角质素聚合物的硫酸角质素、完全不含交联乙酰肝素聚合物的乙酰肝素、完全不含交联硫酸乙酰肝素聚合物的硫酸乙酰肝素或完全不含交联透明质烷聚合物的硫酸透明质烷。 In one aspect of this embodiment, the composition comprises entirely free of cross-linked chondroitin sulfate polymer, chondroitin sulfate, dermatan sulfate entirely free cross dermatan sulfate with polymers, completely free of cross-linked keratin sulfate polymerization transparencies keratan sulfate, heparan entirely free of cross-linked polymer heparan, heparan sulfate entirely free of cross-heparan sulfate-linked polymer or no polymer crosslinked hyaluronan sulfate hyaluronan.

[0064] 本说明书的方面部分地提供了包含一定比例的交联糖胺聚糖聚合物和未交联糖胺聚糖聚合物的水凝胶组合物。 Aspect of the [0064] section of the present specification provides a crosslinked glycosaminoglycan polymer and an uncrosslinked glycosaminoglycan polymer hydrogel composition comprising a certain proportion. 交联和未交联糖胺聚糖聚合物的这种比例也可称为凝胶: 流体比。 Crosslinked and uncrosslinked glycosaminoglycan polymer gels this ratio may also be referred to as: fluid ratio. 在制备本文公开的组合物中任何凝胶:流体比均有用,条件是此比例产生本文公开的改善如本文公开的皮肤状况的组合物。 In preparing the compositions disclosed herein, any of the gel: fluid ratio are used, with the proviso that this ratio produced herein disclosed compositions to improve skin condition as disclosed herein. 本发明组合物中凝胶:流体比的非限制性实例包括100:0、98:2、90:10、75:25、70:30、60:40、50:50、40:60、30:70、25:75、10:90;2:98和0: 100〇 Gel compositions of the present invention: non-limiting examples include fluid than 100: 0,98: 2,90: 10,75: 25,70: 30,60: 40,50: 50, 40: 60, 30: 70,25: 75, 10: 90; 2.: 98 and 0: 100〇

[0065] 在该实施方案的方面,组合物包含交联糖胺聚糖聚合物和未交联糖胺聚糖聚合物,其中凝胶:流体比为(例如)约〇:1〇〇、约1:99、约2:98、约3:97、约4:96、约5:95、约6:94、 约7:93、约8:92、约9:91或约10:90。 [0065] In aspects of this embodiment, the composition comprises a crosslinked glycosaminoglycan polymer and biphenyl uncrosslinked glycosaminoglycan polymer, wherein the gel: fluid ratio (e.g.) about ○: 1〇〇, about 1:99, about 2:98, about 3:97, about 4:96, about 5:95, about 6:94, about 7:93, about 8:92, about 9:91, or about 10:90. 在该实施方案的其它方面,组合物包含交联糖胺聚糖聚合物和未交联糖胺聚糖聚合物,其中凝胶:流体比为(例如)至多1:99、至多2:98、至多3:97、 至多4:96、至多5:95、至多6:94、至多7:93、至多8:92、至多9:91或至多10:90。 In other aspects of this embodiment, the composition comprises a crosslinked glycosaminoglycan polymer and an uncrosslinked glycosaminoglycan polymer, wherein the gel: fluid ratio (e.g.) at most 99, at most 2:98 at most 3:97, at most 4:96, at most 5:95, at most 6:94, at most 7:93, at most 8:92, 9:91 or up to at most 10:90. 在该实施方案的其它方面,组合物包含交联糖胺聚糖聚合物和未交联糖胺聚糖聚合物,其中凝胶:流体比为(例如)约〇:1〇〇至约3:97、约0:100至约5:95或约0:100至约10:90。 In other aspects of this embodiment, the composition comprises a crosslinked glycosaminoglycan polymer and an uncrosslinked glycosaminoglycan polymer, wherein the gel: fluid ratio (e.g.) about ○: 1〇〇 to about 3: 97, from about 0: 100 to about 5:95, or about 0: 100 to about 10:90.

[0066] 在该实施方案的其它方面,组合物包含交联糖胺聚糖聚合物和未交联糖胺聚糖聚合物,其中凝胶:流体比为(例如)约15:85、约20:80、约25:75、约30:70、约35:65、约40:60、 约45:55、约50:50、约55:45、约60:40、约65:35、约70:30、约75:25、约80:20、约85:15、约90: 10、约95:5、约98:2或约100:0。 [0066] In other aspects of this embodiment, the composition comprises a crosslinked glycosaminoglycan polymer and biphenyl uncrosslinked glycosaminoglycan polymer, wherein the gel: fluid ratio (e.g.) about 15:85, about 20 : 80, about 25:75, about 30:70, about 35:65, about 40:60, about 45:55, about 50:50, about 55:45, about 60:40, about 65:35, about 70 : 30, about 75:25, about 80:20, about 85:15, about 90: 10, from about 95: 5, from about 98: 2 or from about 100: 0. 在该实施方案的其它方面,组合物包含交联糖胺聚糖聚合物和未交联糖胺聚糖聚合物,其中凝胶:流体比为(例如)至多15:85、至多20:80、至多25:75、 至多30:70、至多35:65、至多40:60、至多45:55、至多50:50、至多55:45、至多60:40、至多65: 35、至多70:30、至多75:25、至多80:20、至多85:15、至多90:10、至多95:5、至多98:2或至多100:0。 In other aspects of this embodiment, the composition comprises a crosslinked glycosaminoglycan polymer and an uncrosslinked glycosaminoglycan polymer, wherein the gel: fluid ratio (for example) up to 15:85, up to 20:80, up to 25:75, up to 30:70, up to 35:65, up to 40:60, up to 45:55, up to 50:50, up to 55:45, up to 60:40, at most 65: 35, at most 70:30, up to 75:25, up to 80:20, up to 85:15, up to 90:10, at most 95: 5, at most 98: 2 or at most 100: 0. 在该实施方案的其它方面,组合物包含交联糖胺聚糖聚合物和未交联糖胺聚糖聚合物,其中凝胶:流体比为(例如)约10:90至约70:30、约15:85至约70:30、约10:90至约55:45、 约80:20 至约95:5、约90:10 至约100:0、约75:25 至约100:0 或约60:40 至约100:0。 In other aspects of this embodiment, the composition comprises a crosslinked glycosaminoglycan polymer and an uncrosslinked glycosaminoglycan polymer, wherein the gel: fluid ratio (e.g.) about 10:90 to about 70:30, from about 15:85 to about 70:30, from about 10:90 to about 55:45, from about 80:20 to about 95: 5, about 90:10 to about 100: 0, from about 75:25 to about 100: 0 or from about 60:40 to about 100: 0.

[0067] 本文公开的水凝胶组合物可进一步包含在向个体施用所述组合物时提供有益作用的另一种试剂或试剂组合。 [0067] The hydrogel composition disclosed herein may further comprise providing a beneficial effect when the composition is administered to a subject in combination with another agent or agents. 此类有益试剂包括但不限于抗氧化剂、止痒剂、消脂剂、抗疤痕剂、抗炎剂、麻醉剂、抗刺激剂、血管收缩剂、血管扩张剂、抗出血剂(如止血剂或抗纤维溶解蛋白剂)、剥离剂、张紧剂、抗痘剂、色素沉着剂、抗色素沉着剂或保湿剂。 Such benefit agents include but are not limited to, antioxidants, anti-itch agents, anti-cellulite agents, anti-scarring agents, anti-inflammatory agents, anesthetic agents, anti-irritants, vasoconstrictors, vasodilators, anti-bleeding agents (e.g., hemostat or fibrinolytic agent), a release agent, tensioning agents, anti-acne agents, pigmentation agent, an anti-pigmentation agent or a humectant.

[0068] 为了本发明的目的,除非另外说明,否则公式中的“%”定义为重量(S卩,w/w)百分比。 [0068] For purposes of this invention, unless stated otherwise, the formula "%" is defined as the weight (S Jie, w / w) percentages.

[0069] 本发明的方面部分地提供了本文公开的可任选包含麻醉剂的水凝胶组合物。 [0069] aspect of the present invention provide, in part hydrogel composition disclosed herein may optionally contain anesthetic agent. 麻醉剂优选为局部麻醉剂,即引起可逆性局部麻醉和失去伤害感受的麻醉剂,例如氨基酰胺局部麻醉剂和氨基酯局部麻醉剂。 Anesthetic agent is preferably a local anesthetic, i.e. causing reversible local anesthesia and anesthetic nociception loss, such as amino amide local anesthetics and ester local anesthetics amino. 本文公开的组合物中包括的麻醉剂的量是对减轻个体在施用所述组合物后感受到的疼痛有效的量。 The amount of anesthetic agent disclosed herein is included in the composition to alleviate the individual after administration of the composition of an effective amount of pain perceived. 因而,在本说明书中公开的组合物中包括的麻醉剂的量介于按总组合物重量计约0.1%到约5%之间。 Accordingly, the amount of anesthetic agent present in the disclosed compositions include specification interposed by weight of the total composition between about 0.1% to about 5%. 麻醉剂的非限制性实例包括利多卡因、氨布卡因(ambucaine)、阿莫拉酮(arnolanone)、阿米洛卡因(amylocaine)、丁氧普鲁卡因(benoxinate)、苯佐卡因(benzocaine)、贝托卡因(betoxycaine)、苯柳胺酯(biphenamine)、丁昵卡因(bupivacaine)、布他卡因(butacaine)、氨苯丁酯(butamben)、布坦卡因(butanilicaine)、丁胺卡因(butethamine)、丁托西卡因(butoxycaine)、卡铁卡因(carticaine)、氯普鲁卡因(chloroprocaine)、己基苯酸愛康因(cocaethylene)、可卡因(cocaine)、环美卡因(cyclomethycaine)、二丁卡因(dibucaine)、二甲异哇(dimethysoquin)、二甲卡因(dimethocaine)、地昵冬(diperodon)、双环胺(dycyclonine)、 去水芽子减(ecgonidine)、芽子减(ecgonine)、氯乙烧、依替卡因(etidocaine)、β_优卡因(beta-eucaine)、尤普罗辛(euprocin)、非那可明(fenalcomine)、formocaine、海克卡因(hexylcaine)、轻丁卡因(hydroxy teracaine)、对 Non-limiting examples of anesthetics include lidocaine, dibucaine ammonia (ambucaine), Amora ketone (arnolanone), because Ami Loka (amylocaine), oxybuprocaine (benoxinate), benzocaine (benzocaine), because Beituo Ka (betoxycaine), Liu amine phenyl ester (biphenamine), D LIMITED tetracaine (bupivacaine), butacaine (butacaine), butamben (butamben), because Butan Ka (butanilicaine ), tetracaine butylamine (butethamine), Dingtuo by Sika (butoxycaine), iron tetracaine card (carticaine), chloroprocaine (chloroprocaine), benzene hexanoic acid Aikang by (cocaethylene), cocaine (cocaine) , cyclomethycaine (cyclomethycaine), dibucaine (dibucaine), dimethyl iso-wow (dimethysoquin), dimethyl tetracaine (dimethocaine), Nick the winter (diperodon), bicyclic amines (dycyclonine), to the water sprouts Save (ecgonidine), Save sprouts (ecgonine), burning chloride, etidocaine (etidocaine), β_ preferably lidocaine (beta-eucaine), Youpu Rossing (euprocin), it can be non-Ming (fenalcomine), formocaine, hexylcaine (hexylcaine), light tetracaine (hydroxy teracaine), of 基苯甲酸异丁酯、甲磺酸亮氨卡因(leucinocaine mesylate)、左沙屈尔(Ievoxadrol)、利多卡因、甲昵卡因(mepivacaine)、 美普卡因(meprylcaine)、美布卡因(metabutoxycaine)、氯甲烧、麦替卡因(myrtecaine)、 纳伊卡因(naepaine)、奥他卡因(octacaine)、奥索卡因(orthocaine)、轻乙卡因(oxethazaine)、对乙氧卡因(parethoxycaine)、非那卡因(phenacaine)、苯酸、昵罗卡因(piperocaine)、匹多卡因(piridocaine)、聚多卡醇(polidocanol)、丙吗卡因(pramoxine)、丙胺卡因(prilocaine)、普鲁卡因(procaine)、丙泮卡因(propanocaine)、丙美卡因(proparacaine)、丙脈卡因(propipocaine)、丙氧卡因(propoxycaine)、假可卡因(psuedococaine)、卩比略卡因(pyrrocaine)、罗昵卡因(ropi vacaine)、水杨醇、丁卡因(tetracaine)、托利卡因(tolycaine)、三甲卡因(trimecaine)、佐拉敏(zolamine)、其组合及其盐。 Acid isobutyl ester, methanesulfonic acid leucine lidocaine (leucinocaine mesylate), the left flexor sand Seoul (Ievoxadrol), lidocaine, tetracaine A nickname (mepivacaine), meprylcaine (meprylcaine), United States dibucaine by (metabutoxycaine), chloroformates burning, wheat articaine (myrtecaine), Na Yika because (naepaine), Austrian he tetracaine (octacaine), by Ozark (orthocaine), tetracaine light b (oxethazaine), p-ethoxy tetracaine (parethoxycaine), non-phenacaine (phenacaine), benzoic acid, by Nick Roca (piperocaine), horses lidocaine (piridocaine), polidocanol (polidocanol), pramoxine (pramoxine), propylamine Cain (prilocaine), procaine (procaine), propylene Pan Cain (propanocaine), proparacaine (proparacaine), propylene pulse Cain (propipocaine), propoxycaine (propoxycaine), fake cocaine (psuedococaine ), Jie slightly than bupivacaine (pyrrocaine), Nick Luo ropivacaine (ropi vacaine), salicyl alcohol, tetracaine (tetracaine), because Tolka (tolycaine), trimecaine (trimecaine), Zuo Lamin ( zolamine), combinations thereof and salts thereof. 氨基酯局部麻醉剂包括普鲁卡因、氯普鲁卡因、可卡因、环美卡因、cimethocaine (拉罗卡因(Iarocaine))、丙氧卡因、普鲁卡因(奴佛卡因(novocaine))、丙美卡因、丁卡因(阿美索卡因(amethocaine))。 Amino ester local anesthetics include procaine, chloroprocaine, cocaine, cyclomethycaine, cimethocaine (LaRocca by (Iarocaine)), propoxycaine, procaine (novocaine (Novocaine )), proparacaine, tetracaine (amethocaine (amethocaine)). 氨基酰胺局部麻醉药的非限制性实例包括阿替卡因(articaine)、丁昵卡因、辛可卡因(cinchocaine)(二丁卡因)、依替卡因、左布比卡因(Ievobupivacaine)、利多卡因(利诺卡因(Iignocaine))、甲昵卡因、昵罗卡因、丙胺卡因、 罗昵卡因和三甲卡因。 Non-limiting examples of amino amide local anesthetics include articaine (articaine), D Nick tetracaine, dibucaine (cinchocaine) (dibucaine), etidocaine, levobupivacaine (Ievobupivacaine), lidocaine (lignocaine (Iignocaine)), A Nick Cain, Nick Rocca, prilocaine, and ropivacaine Luo Nick trimecaine. 本文公开的组合物可包含单种麻醉剂或多种麻醉剂。 The compositions disclosed herein can comprise single or multiple anesthetic anesthetic agents. 组合局部麻醉药的非限制性实例为利多卡因/丙胺卡因(EMLA)。 Non-limiting examples of combinations of local anesthetics lidocaine / prilocaine (EMLA).

[0070] 因此在一个实施方案中,本文公开的组合物包含麻醉剂及其盐。 [0070] Thus, in one embodiment, the herein disclosed compositions comprising anesthetic agents and salts thereof. 在该实施方案的方面,本文公开的组合物包含氨基酰胺局部麻醉剂及其盐或氨基酯局部麻醉剂及其盐。 In aspects of this embodiment, the compositions disclosed herein comprise an amino amide local anesthetic and salts or aminoester local anesthetic and salts thereof. 在该实施方案的其它方面,本文公开的组合物包含普鲁卡因、氯普鲁卡因、可卡因、环美卡因、 cimethocaine、丙氧卡因、普鲁卡因、丙美卡因、丁卡因或其盐或其任何组合。 In other aspects of this embodiment, the compositions disclosed herein comprise procaine, chloroprocaine, cocaine, cyclomethycaine, cimethocaine, propoxycaine, procaine, proparacaine, D tetracaine or a salt thereof, or any combination thereof. 在该实施方案的其它方面,本文公开的组合物包含阿替卡因、丁昵卡因、辛可卡因、依替卡因、左布比卡因、利多卡因、甲昵卡因、昵罗卡因、丙胺卡因、罗昵卡因、三甲卡因或其盐或其任何组合。 In other aspects of this embodiment, the composition disclosed herein comprises articaine, tetracaine Nick butoxy, cinchocaine, etidocaine, levobupivacaine, lidocaine, tetracaine A Nick, Nick Rocca , prilocaine, ropivacaine Luo Nick, trimecaine or salts or any combination thereof. 在该实施方案的另外其它方面,本文公开的组合物包含利多卡因/丙胺卡因组合。 In still other aspects of this embodiment, the compositions disclosed herein comprise lidocaine / prilocaine combination.

[0071] 在该实施方案的其它方面,本文公开的组合物包含按总组合物的重量计,量为(例如)约0.1 %、约0.2%、约0.3%、约0.4%、约0.5%、约0.6%、约0.7%、约0.8%、约0.9%、约1.0%、约2.0%、约3.0%、约4.0%、约5.0%、约6.0%、约7.0%、约8.0%、约9.0%或约10% 的麻醉剂。 [0071] In other aspects of this embodiment, the herein disclosed composition comprises, by weight of the total composition, an amount of (e.g.) about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1.0%, about 2.0%, about 3.0%, about 4.0%, about 5.0%, about 6.0%, about 7.0%, about 8.0%, about 9.0 % or about 10% of the anesthetic. 在另外其它方面,本文公开的组合物包含按总组合物的重量计,量为(例如)至少0.1 %、至少0.2%、至少0.3%、至少0.4%、至少0.5%、至少0.6%、至少0.7 %、至少0.8%、 至少0.9 %、至少1.0 %、至少2.0 %、至少3.0 %、至少4.0 %、至少5.0 %、至少6.0 %、至少7.0 %、至少8.0 %、至少9.0 %或至少10 %的麻醉剂。 In still other aspects, the herein disclosed composition comprises the total composition weight, an amount of (e.g.) at least 0.1%, at least 0.2%, at least 0.3%, at least 0.4%, at least 0.5%, at least 0.6%, at least 0.7 %, at least 0.8%, at least 0.9%, at least 1.0%, at least 2.0%, at least 3.0%, at least 4.0%, at least 5.0%, at least 6.0%, at least 7.0%, at least 8.0%, at least 9.0% or at least 10% of the Anesthetic. 在还有其它方面,本文公开的组合物包含按总组合物的重量计,量为(例如)至多0.1 %、至多0.2%、至多0.3%、至多0.4%、至多0.5%、至多0.6%、至多0.7%、至多0.8%、至多0.9%、至多1.0%、至多2.0%、至多3.0%、 至多4.0%、至多5.0%、至多6.0%、至多7.0%、至多8.0%、至多9.0%或至多10%的麻醉剂。 In yet other aspects, the herein disclosed composition comprises by weight of the total composition, an amount of (e.g.) at most 0.1%, at most 0.2%, at most 0.3%, at most 0.4%, at most 0.5%, at most 0.6%, at most 0.7% up to 0.8%, up to 0.9%, up to 1.0%, at most 2.0%, at most 3.0%, at most 4.0%, at most 5.0%, at most 6.0%, at most 7.0%, at most 8.0%, at most 9.0% or at most 10% the anesthetic. 在其它方面,本文公开的组合物包含按总组合物的重量计,量为(例如)约〇. 1 %至约0.5%、约0.1 % 至约1.0%、约0.1 %至约2.0%、约0.1 % 至约3.0%、约0.1 %至约4.0%、约0.1 %至约5.0%、约0.2%至约0.9%、约0.2%至约1.0%、约0.2%至约2.0%、约0.5%至约1.0%或约0.5%至约2.0%的麻醉剂。 In other aspects, the herein disclosed composition comprises the total composition weight, an amount of (e.g.) about billion. 1% to about 0.5%, about 0.1% to about 1.0%, about 0.1% to about 2.0%, about 0.1% to about 3.0%, about 0.1% to about 4.0%, about 0.1% to about 5.0%, about 0.2% to about 0.9%, about 0.2% to about 1.0%, about 0.2% to about 2.0%, about 0.5% to about 1.0%, or from about 0.5% to about 2.0% of the anesthetic.

[0072] 在另一实施方案中,本文公开的组合物不包含麻醉剂。 [0072] In another embodiment, the herein disclosed composition does not contain anesthetic agent.

[0073] 在本发明的一个方面,提供了可注射真皮填充剂,其包含聚合物,例如糖胺聚糖聚合物,例如透明质酸聚合物,例如至少一部分交联的透明质酸,和添加剂或与所述聚合物组合的有益试剂。 [0073] In one aspect of the present invention, there is provided an injectable dermal filler comprising a polymer, glycosaminoglycan polymer, for example, a polymer such as hyaluronic acid, for example, at least a portion crosslinked hyaluronic acid, and additives or in combination with the polymer benefit agent.

[0074] 与所述聚合物组合的有益试剂包含维生素,例如维生素C。 [0074] combined with the polymer benefit agent comprises vitamin, such as vitamin C. 适合形式的维生素C的非限制性实例包括抗坏血酸和抗坏血酸的钠盐、钾盐和钙盐、抗坏血酸与长链脂肪酸的脂溶性酯(抗坏血酸棕榈酸酯或抗坏血酸硬脂酸酯)、抗坏血酸磷酸酯镁(MAP)、抗坏血酸磷酸钠(SAP)和抗坏血酸2-葡萄糖苷(AA2G™)、抗坏血酸磷酸钠(AA2P)、抗坏血酸硫酸二钠和抗坏血酸3-氨丙基磷酸酯(维生食物)。 Non-limiting examples of suitable form of vitamin C and ascorbic acid include ascorbic acid, sodium, potassium and calcium salts of ascorbic acid with long chain fatty acids, fat-soluble ester (ascorbyl palmitate or ascorbyl stearate), magnesium ascorbyl phosphate (MAP), sodium ascorbyl phosphate (the SAP) and ascorbic acid 2-glucoside (AA2G ™), sodium ascorbyl phosphate (AA2P), disodium ascorbyl sulfate, and 3-aminopropyl ascorbyl phosphate (vitamin food).

[0075] 在一个特别有利的实施方案中,有益试剂与所述聚合物共价结合。 [0075] In a particularly advantageous embodiment, the beneficial agent is covalently bound to the polymer. 例如,有益试剂可为维生素C或维生素C衍生物,其与所述聚合物共价结合并且按总组合物的重量计,在组合物中存在的量介于约〇.04%至约5.0%之间,例如按总组合物的重量计介于约0.1 %至约4.0 %之间,例如按总组合物的重量计介于约0.2 %至约2.0 %之间。 For example, the beneficial agent may be vitamin C or a vitamin C derivative, which is covalently bound to the polymer and the total weight of the composition, it is present in the composition in an amount ranging from about 5.0% to about 〇.04% between, for example, by weight of the total composition is between about 0.1% to about 4.0%, for example, by weight of the total composition is between about 0.2% to about 2.0%. 在一个实施方案中,本文公开的组合物中包括的维生素C的量按总组合物的重量计介于约0.3%至约1.2%之间。 In one embodiment, the amount of vitamin C in the herein disclosed composition comprises by weight of the total composition is between about 0.3% to about 1.2%.

[0076] 优选地,与所述组合物共价结合的维生素C包括下列至少一种:抗坏血酸、L-抗坏血酸、L-抗坏血酸2-硫酸酯(AA-2S)和L-抗坏血酸2-磷酸酯(AA-2P)、抗坏血酸2-0-葡萄糖苷(AA-2G)、6-0_酰基-2-0-aD-吡喃葡萄糖基-L-抗坏血酸(6-酰基-AA-2G)、(抗坏血酸3-氨丙基磷酸酯、抗坏血酸棕榈酸酯)、其衍生物和组合。 [0076] Preferably, the covalent binding of the vitamin C composition comprising at least one of the following: ascorbic acid, L- ascorbic acid, L- ascorbic acid 2-sulfate (AA-2S) and L- ascorbic acid 2-phosphate ( AA-2P), ascorbic acid glucoside 2-0- (AA-2G), 6-0_ acyl -2-0-aD- -L- glucopyranosyl-ascorbic acid (6-acyl -AA-2G), (ascorbic acid 3-aminopropyl phosphate, ascorbyl palmitate), derivatives thereof, and combinations thereof. 本文公开的组合物可包含单种维生素C试剂或多种维生素C试剂。 The compositions disclosed herein may comprise a single agent or more of vitamin C, vitamin C agents.

[0077] 在本发明的另一实施方案中,提供了真皮填充剂,其中透明质酸与BDDE交联。 [0077] In another embodiment of the present invention, there is provided a dermal filler, wherein the crosslinked hyaluronic acid BDDE. 在这个实施方案中,结合度可介于约3mol %和约IOmol %之间,约15mol %到约40mol %之间。 In this embodiment, the degree of binding may be between about 3mol% and about IOmol%, between about 15mol% to about 40mol%.

[0078] 在一些实施方案中,真皮填充剂具有持续的生物利用率。 [0078] In some embodiments, the dermal filler has sustained bioavailability. 例如,提供了在引入人类皮肤中时,向人类有效释放抗坏血酸或其它维生素至少约1个月或长达约20个月或更长时间的真皮填充剂。 For example, provided in the introduction into the human skin, the effective release of ascorbic acid or other vitamins at least about one month, or up to about 20 months or longer to human dermal fillers.

[0079] 本发明的方面部分地提供了本文公开的显示复数模量、弹性模量、粘性模量和tan 如勺水凝胶组合物。 [0079] aspect of the present invention provide, in part display complex modulus disclosed herein, elastic modulus, viscous modulus and tan scoop hydrogel composition. 当施加力(应力、变形)时,本文公开的组合物具粘弹性,因为组合物具有弹性组分(固体状例如交联糖胺聚糖聚合物)和粘性组分(液体状例如未交联的糖胺聚糖聚合物或载体相)。 When a force (stress, deformation), a composition disclosed herein has viscoelastic, because the composition has an elastic component (e.g. a solid crosslinked glycosaminoglycan polymer) and the viscous component (e.g. liquid uncrosslinked the glycosaminoglycan polymer or carrier). 描述该性质的流变学属性是复数模量(G*),其定义组合物对变形的总抗性。 The rheological properties of the described properties are complex modulus (G *), the total resistance to deformation defined composition. 复数模量是具有实部和虚部的复数:G+iG^iG'G*的绝对值为Abs (G*) =Sqrt (G/2+G 〃2)。 The complex modulus is a complex real and imaginary part: G + iG iG'G ^ * of the absolute value Abs (G *) = Sqrt (G / 2 + G 〃2). 可将复数模量定义为弹性模量(G')和粘性模量(G”)的总和。Falcone等,Temporary Polysaccharide Dermal Fillers:A Model for Persistence Based on Physical Properties,De;rmatol Surg.35 (8) :1238-1243 (2009) ;Tezel,同上,2008;Kablik,同上, 2009; Beasley,同上,2009;其各自据此通过引用整体并入。 Complex modulus may be defined as the elastic modulus (G ') and viscous modulus (G ") sum .Falcone like, Temporary Polysaccharide Dermal Fillers: A Model for Persistence Based on Physical Properties, De; rmatol Surg.35 (8 ): 1238-1243 (2009); Tezel, ibid., 2008; Kablik, ibid., 2009; Beasley, supra, 2009; each of which is hereby incorporated by reference.

[0080] 弹性模量或弹性模数指水凝胶材料抵抗变形的能力,或相反地,在对其施加力时物体对非永久性变形的倾向。 [0080] The modulus of elasticity or elastic modulus refers to the ability to resist deformation of the hydrogel material, or, conversely, the tendency of the non-permanent deformation of the object when a force is applied to them. 弹性模量表征了组合物的牢固性,并且因为它描述了来自组合物运动的能量储存,所以也被称为储能模量。 It characterized a solid elastic modulus of the composition, and because it describes the movement of energy from the storage composition, is also known as the storage modulus. 弹性模量描述了弹性和强度之间的相互作用(G' =应力/应变),并因此提供了对组合物硬度或柔软度的定量测量。 It describes elastic modulus and strength of the interaction between the elastic (G '= stress / strain), and therefore provides a quantitative measure of the hardness or softness of the composition of. 将物体的弹性模量定义为在弹性变形区域中其应力-应变曲线的斜率:λ =应力/应变,其中λ是帕斯卡定理(Pascal's)中的弹性模量;应力是引起变形的力除以施加力的面积;并且应变是应力引起的变化与物体初始状态之比。 The elastic modulus of the defined object as the elastic deformation region stress - strain curve of slope: λ = stress / strain, where [lambda] is the first theorem (Pascal's) modulus of elasticity in; the stress is the force caused by the deformation divided by the applied area power; and the strain is the ratio of stress-induced changes in the initial state of the object. 虽然取决于施加力的速度,但组合物越硬,具有的弹性模量越高并且将花更大的力使材料在指定距离变形,例如注射。 Although the applied force depends on the speed, but the harder the composition, having the higher modulus of elasticity and spend a greater force at a specified distance of the material deformation, for example, injection. 说明如何测量应力,包括方向,允许定义许多类型的弹性模量。 How to measure stress, including direction, allowing definition of the elastic modulus of many types. 3个主要弹性模量为拉伸模量、剪切模量和体积模量。 3 main elastic modulus of tensile modulus, shear modulus and bulk modulus.

[0081] 粘性模量也称为损耗模量,因为它描述了随粘性耗散而损耗的能量。 [0081] also referred to as viscous modulus loss modulus, because it describes the energy rises with viscous dissipation loss. TanS为粘性模量和弹性模量之比,tan5 = G”/G' jalcone,同上,2009。对于在本说明书中公开的tan5 值,tanS由在IHz频率下的动态模量得到。更低的tanS对应于更硬的、更坚固或更有弹性的组合物。 TanS viscous modulus and the elastic modulus ratio, tan5 = G "/ G 'jalcone, supra, 2009 for tan5 values ​​disclosed in the present specification, TANS obtained by the dynamic modulus at IHz frequency. Lower tanS corresponds to a harder, more rigid or more flexible compositions.

[0082] 在另一实施方案中,本文公开的水凝胶组合物显示出弹性模量。 [0082] In another embodiment, disclosed herein hydrogel composition exhibits an elastic modulus. 在该实施方案的方面,水凝胶组合物显示出(例如)约25Pa、约50Pa、约75Pa、约IOOPa、约125Pa、约150Pa、约175Pa、约200Pa、约250Pa、约300Pa、约350Pa、约400Pa、约450Pa、约500Pa、约550Pa、约600Pa、约650Pa、约700Pa、约750Pa、约800Pa、约850Pa、约900Pa、约950Pa、约l,000Pa、约1, 200Pa、约I,300Pa、约I,400Pa、约I,500Pa、约I,600Pa、约1700Pa、约1800Pa、约1900Pa、约2, OOOPa、约2,100Pa、约2,200Pa、约2,300Pa、约2,400Pa或约2,500Pa的弹性模量。 In aspects of this embodiment, a hydrogel composition exhibits (e.g.) about 25Pa, about 50Pa, about 75Pa, about IOOPa, about 125Pa, 150Pa about, about 175 Pa, about 200Pa, 250Pa about, about 300Pa, about 350 Pa, about 400Pa, about 450Pa, about 500Pa, about 550Pa, about 600Pa, about 650Pa, about 700Pa, about 750Pa, about 800Pa, about 850Pa, about 900Pa, about 950Pa, about l, 000Pa, from about 1, 200Pa, about I, 300Pa , about I, 400Pa, about I, 500Pa, about I, 600Pa, about 1700Pa, about 1800 Pa, from about 1900 Pa, about 2, OOOPa, about 2,100Pa, about 2,200Pa, about 2,300Pa, or from about about 2,400Pa 2,500Pa elastic modulus. 在该实施方案的其它方面,水凝胶组合物显示出(例如)至少25Pa、至少50Pa、至少75Pa、至少lOOPa、至少125Pa、至少150Pa、至少175Pa、至少200Pa、至少250Pa、至少300Pa、至少350Pa、至少400Pa、至少450Pa、至少500Pa、至少550Pa、至少600Pa、至少650Pa、至少700Pa、至少750Pa、 至少800Pa、至少850Pa、至少900Pa、至少950Pa、至少I,000Pa、至少I,200Pa、至少I,300Pa、 至少l,400Pa、至少l,500Pa、至少l,600Pa、至少1700Pa、至少1800Pa、至少1900Pa、至少2, 000Pa、至少2,100Pa、至少2,200Pa、至少2,300Pa、至少2,400Pa或至少2,500Pa的弹性模量。 In other aspects of this embodiment, a hydrogel composition exhibits (e.g.) at least 25Pa, at least 50Pa, at least 75Pa, at least Loopa, at least 125Pa, 150Pa at least, at least 175 Pa, at least 200Pa, 250Pa at least, at least 300Pa, 350Pa least at least of 400 Pa, at least 450Pa, at least 500Pa, at least 550 Pa, at least 600Pa, at least 650Pa, at least 700Pa, at least 750 Pa, at least 800 Pa, at least 850 Pa, at least 900Pa, at least 950Pa, at least I, 000 Pa, at least I, 200Pa, at least I, 300Pa, at least l, 400Pa, at least l, 500Pa, at least l, 600Pa, at least 1700Pa, at least 1800Pa, at least 1900Pa, at least 2, 000Pa, at least 2,100Pa, at least 2,200Pa, at least 2,300Pa, at least 2,400Pa or the elastic modulus of at least 2,500Pa. 在该实施方案的另外其它方面,水凝胶组合物显示出(例如)至多25Pa、至多50Pa、至多75Pa、至多100Pa、至多125Pa、至多150Pa、至多175Pa、至多200Pa、至多250Pa、至多300Pa、至多350Pa、至多400Pa、至多450Pa、至多500Pa、至多550Pa、至多600Pa、至多650Pa、至多700Pa、至多750Pa、至多800Pa、至多850Pa、至多900Pa、至多950Pa、至多l,000Pa、至多1, 200Pa、至多I,300Pa、至多I,400Pa、至多I,500Pa或至多I,600Pa的弹性模量。 In still other aspects of this embodiment, a hydrogel composition exhibits (e.g.) at most 25Pa, at most 50Pa, at most 75Pa, at most 100 Pa, at most 125Pa, 150Pa at most, at most 175 Pa, at most 200Pa, 250Pa at most, at most 300Pa, most 350Pa, at most 400Pa, at most 450Pa, at most 500Pa, at most 550Pa, at most 600Pa, at most 650Pa, at most 700Pa, at most 750Pa, at most 800Pa, at most 850Pa, at most 900Pa, at most 950Pa, up to l, 000Pa, at most 1, 200Pa, up to I , 300Pa, at most I, 400Pa, at most I, or up to 500Pa I, the modulus of elasticity of 600Pa. 该实施方案的另外其它方面,水凝胶组合物显示出(例如)约25Pa至约150Pa、约25Pa至约300Pa、约25Pa至约500Pa、约25Pa 至约800Pa、约125Pa 至约300Pa、约125Pa 至约500Pa、约125Pa 至约800Pa、约500Pa至约I,600Pa、约600Pa至约I,600Pa、约700Pa至约I,600Pa、约800Pa至约I,600Pa、约900Pa至约I,600Pa、约I,OOOPa至约I,600Pa、约I,IOOPa至约I,600Pa、约I,200Pa至约1, 600Pa、约500Pa至约2,500Pa、约I,OOOPa至约2,500Pa、约I,500Pa至约2,500Pa、约2,OOOPa 至约2,500Pa、约I,300Pa至约I,600Pa、约I,400Pa至约I,700Pa、约I,500Pa至约I,800Pa、约I,600Pa至约I,900Pa、约I,700Pa至约2,OOOPa、约I,800Pa至约2,IOOPa、约I,900Pa至约2, 200Pa、约2,OOOPa至约2,300Pa、约2,IOOPa至约2,400Pa或约2,200Pa至约2,500Pa的弹性模量。 Still other aspects, the hydrogel composition exhibits this embodiment (e.g.) from about 25Pa to about 150Pa, about 25Pa to about 300Pa, about 25Pa to about 500Pa, about 25Pa to about 800 Pa, from about 125Pa to about 300Pa, 125Pa to about to about 500Pa, from about 125Pa to about 800Pa, from about 500Pa to about I, 600Pa, from about 600Pa to about I, 600Pa, from about 700Pa to about I, 600Pa, from about 800Pa to about I, 600Pa, from about 900Pa to about I, 600Pa, about I, OOOPa about I, 600Pa, about I, IOOPa about I, 600Pa, about I, 200Pa to about 1, 600Pa, from about 500Pa to about 2,500Pa, about I, OOOPa about 2,500Pa, about I , 500Pa to about 2,500Pa, about 2, OOOPa about 2,500Pa, about I, 300Pa to about I, 600Pa, about I, 400Pa to about I, 700Pa, about I, 500Pa to about I, 800Pa, about I , 600Pa to about I, 900Pa, about I, 700Pa to about 2, OOOPa, about I, 800Pa to about 2, IOOPa, about I, 900Pa to about 2, 200Pa, about 2, OOOPa about 2,300Pa, about 2 , IOOPa to the elastic modulus of about 2,400Pa or about to about 2,500Pa of 2,200Pa.

[0083] 在另一实施方案中,本文公开的水凝胶组合物显示出粘性模量。 [0083] In another embodiment, disclosed herein hydrogel composition exhibits a viscous modulus. 在该实施方案的方面,水凝胶组合物显示出(例如)约IOPa、约20Pa、约30Pa、约40Pa、约50Pa、约60Pa、约70Pa、约80Pa、约90Pa、约100Pa、约150Pa、约200Pa、约250Pa、约300Pa、约350Pa、约400Pa、约450Pa、约500Pa、约550Pa、约600Pa、约650Pa或约700Pa的粘性模量。 In aspects of this embodiment, a hydrogel composition exhibits (e.g.) about IOPa, about 20Pa, about 30 Pa, about of 40 Pa, about 50Pa, about 60Pa, about 70Pa, about 80Pa, about 90Pa, about 100 Pa, about 150Pa, about 200Pa, about 250Pa, about 300Pa, about 350Pa, about 400Pa, about 450Pa, about 500Pa, about 550Pa, about 600Pa, the viscous modulus of from about 700Pa to about 650Pa, or. 在该实施方案的其它方面,水凝胶组合物显示出(例如)至多l〇Pa、至多20Pa、至多30Pa、至多40Pa、至多50Pa、至多60Pa、至多70Pa、至多80Pa、至多90Pa、至多lOOPa、至多150Pa、至多200Pa、至多250Pa、至多300Pa、至多350Pa、至多400Pa、至多450Pa、至多500Pa、至多550Pa、至多600Pa、至多650Pa 或至多700Pa的粘性模量。 In other aspects of this embodiment, a hydrogel composition exhibits (e.g.) l〇Pa up to, at most 20Pa, at most 30 Pa, at most of 40 Pa, at most 50Pa, at most 60Pa, at most 70Pa, at most 80Pa, at most 90Pa, at most Loopa, at most 150Pa, at most 200Pa, at most 250Pa, at most 300Pa, at most 350Pa, at most 400Pa, at most 450Pa, at most 500Pa, at most 550Pa, at most 600Pa, the viscous modulus of at most 650Pa or at most 700Pa. 在该实施方案的另外其它方面,水凝胶组合物显示出(例如)约IOPa 至约30Pa、约IOPa至约50Pa、约IOPa至约lOOPa、约IOPa至约150Pa、约70Pa至约lOOPa、约50Pa 至约350Pa、约150Pa 至约450Pa、约250Pa 至约550Pa、约350Pa 至约700Pa、约50Pa 至约150Pa、约IOOPa 至约200Pa、约150Pa 至约250Pa、约200Pa 至约300Pa、约250Pa 至约350Pa、约300Pa 至约400Pa、约350Pa 至约450Pa、约400Pa 至约500Pa、约450Pa 至约550Pa、约500Pa 至约600Pa、约550Pa至约650Pa或约600Pa至约700Pa的粘性模量。 In still other aspects of this embodiment, a hydrogel composition exhibits (e.g.) about IOPa to about 30 Pa, about 50Pa to about IOPa, about IOPa about Loopa, about IOPa to about 150Pa, about 70Pa to about Loopa, about 50Pa to about 350Pa, from about 150Pa to about 450Pa, from about 250Pa to about 550Pa, from about 350 Pa to about 700Pa, about 50Pa to about 150Pa, about IOOPa to about 200Pa, from about 150Pa to about 250Pa, from about 200Pa to about 300Pa, about 250Pa to about 350Pa, from about 300Pa to about 400Pa, from about 350 Pa to about 450Pa, from about 400Pa to about 500Pa, from about 450Pa to about 550Pa, from about 500Pa to about 600Pa, the viscous modulus of about 550 Pa, or from about 600Pa to about 650Pa to about 700Pa.

[0084] 在另一实施方案中,本文公开的水凝胶组合物显示出tanS。 [0084] In another embodiment, disclosed herein hydrogel composition exhibits tanS. 在该实施方案的方面, 水凝胶组合物显示出(例如)约0.1、约0.2、约0.3、约0.4、约0.5、约0.6、约0.7、约0.8、约0.9、约1.0、约1.1、约1.2、约1.3、约1.4、约1.5、约1.6、约1.7、约1.8、约1.9、约2.0、约2.1、 约2.2、约2.3、约2.4或约2.5的tanS。 In aspects of this embodiment, a hydrogel composition exhibits (e.g.) about 0.1, about 0.2, about 0.3, about 0.4, about 0.5, about 0.6, about 0.7, about 0.8, about 0.9, about 1.0, about 1.1, about 1.2, about 1.3, about 1.4, about 1.5, about 1.6, about 1.7, about 1.8, about 1.9, about 2.0, about 2.1, about 2.2, about 2.3, about 2.4, or about 2.5 tanS. 在该实施方案的其它方面,水凝胶组合物显示出(例如)至多0.1、至多0.2、至多0.3、至多0.4、至多0.5、至多0.6、至多0.7、至多0.8、至多0.9、至多1.0、至多1.1、至多1.2、至多1.3、至多1.4、至多1.5、至多1.6、至多1.7、至多1.8、至多1.9、至多2.0、至多2.1、至多2.2、至多2.3、至多2.4或至多2.5的tanS。 In other aspects of this embodiment, a hydrogel composition exhibits (e.g.) at most 0.1, at most 0.2, at most 0.3, at most 0.4, at most 0.5, at most 0.6, at most 0.7, at most 0.8, at most 0.9, at most 1.0, at most 1.1 , at most 1.2, at most 1.3, at most 1.4, at most 1.5, at most 1.6, at most 1.7, at most 1.8, at most 1.9, at most 2.0, at most 2.1, at most 2.2, at most 2.3, tanS at most 2.4 or at most 2.5. 在该实施方案的另外其它方面,水凝胶组合物显示出(例如)约〇. 1至约〇. 3、约0.3至约0.5、约0.5至约0.8、约1.1 至约1.4、约1.4至约1.7、约0.3至约0.6、约0.1至约0.5、约0.5至约0.9、约0.1至约0.6、约0.1至约1.0、约0.5至约1.5、约1.0至约2.0或约1.5至约2.5的tanS。 In still other aspects of this embodiment, a hydrogel composition exhibits (e.g.) square to about 1 billion to about 3, from about 0.3 to about 0.5, from about 0.5 to about 0.8, from about 1.1 to about 1.4, from about 1.4 to about 1.7, about 0.3 to about 0.6, from about 0.1 to about 0.5, from about 0.5 to about 0.9, from about 0.1 to about 0.6, from about 0.1 to about 1.0, from about 0.5 to about 1.5, from about 1.0 to about 2.0, or from about 1.5 to about 2.5 the tanS.

[0085] 本说明书的方面部分地提供了本文公开的具有透明性和/或半透明性的水凝胶组合物。 Aspect of the [0085] present specification provide, in part transparent and / or translucent hydrogels having compositions disclosed herein. 光学透明性是允许可见光通过材料的物理性质,而半透明性(也称为半透明或半透明度)仅允许光漫射通过。 Optical transparency that allows visible light to pass physical properties of the material, while the translucency (also referred to as a translucent or semi-transparency) only allows through light diffusion. 相对的性质是不透明性。 Properties of the relative opacity is. 透明材料清澈,而半透明材料不能清楚地看透。 Clear transparent material, a translucent material and can not be clearly discerned. 本文公开的水凝胶优选为光学透明或至少半透明。 Disclosed herein, a hydrogel is preferably optically transparent or at least translucent.

[0086] 在一个实施方案中,本文公开的水凝胶组合物光学透明。 [0086] In one embodiment, disclosed herein, optically clear hydrogel composition. 在该实施方案的方面,水凝胶组合物漫射传导(例如)约75 %的光、约80 %的光、约85 %的光、约90 %的光、约95 %的光或约100 %的光。 In aspects of this embodiment, the diffusion conductive hydrogel composition (e.g.,) about 75% of light, a light of about 80%, about 85% of light, a light of about 90%, or about 95 to about 100% of the light % of the light. 在该实施方案的其它方面,水凝胶组合物漫射传导(例如)至少75 %的光、至少80%的光、至少85%的光、至少90%的光或至少95%的光。 In other aspects of this embodiment, the diffusion conductive hydrogel composition (e.g.) at least 75% of the light, the light of at least 80%, at least 85% of light, at least 90% or at least 95% of the optical light. 在该实施方案的另外其它方面,水凝胶组合物漫射传导(例如)约75 %至约100 %的光、约80 %至约100 %的光、约85%至约100 %的光、约90%至约100%的光或约95%至约100 %的光。 In still other aspects, the diffusion conductive hydrogel composition (e.g.,) about 75% to about 100% of the light, from about 80% to about 100% of the light, from about 85% to about 100% of the light of this embodiment, from about 90% to about 100% of the light, or from about 95% to about 100% of the light. 在一个实施方案中, 本文公开的水凝胶组合物光学透明并且传导100%的可见光。 In one embodiment, the herein disclosed hydrogel composition and an optically transparent conductive visible light of 100%.

[0087] 可通过将水凝胶磨成颗粒进一步加工本文公开的水凝胶组合物并且任选与载体相例如水或盐水溶液混合以形成可注射或局部物质,如溶液、油、洗液、凝胶、软膏、乳膏、浆料、油膏或糊剂。 [0087] The hydrogel can be ground into particles by the process herein further disclosed hydrogel composition and optionally mixed with a carrier, for example, aqueous or saline solution to form an injectable or topical substances, such as solutions, oils, lotions, gel, ointment, cream, paste, ointment or paste. 因而,公开的水凝胶组合物可为单相或多相组合物。 Thus, the disclosed hydrogel composition may be a single phase or multiphase composition. 可将水凝胶碾磨成直径约I Oym至约100 Ομπι的粒度,例如约15μηι至约3 Ομπι、约5 Ομπι至约7 5μηι、约1 ΟΟμπι至约15 Oym、 约200μηι 至约300μηι、约450μηι至约550μηι、约600μηι 至约700μηι、约750μηι至约850μηι或约900μηι 至约1,000μπι。 The hydrogel may be milled to a diameter of about I Oym about 100 Ομπι particle size, for example from about 15μηι to about 3 Ομπι, about 5 Ομπι to about 7 5μηι, from about 1 ΟΟμπι to about 15 Oym, from about to about 200μηι 300μηι, about 450μηι about 550μηι, about 600μηι about 700μηι, or about 750μηι about 850μηι about 900μηι about 1,000μπι.

[0088] 本发明的方面部分地提供了本文公开的可注射的组合物。 [0088] aspect of the present invention provide, in part injectable compositions disclosed herein. 如本文所使用,术语“可注射”指材料具有使用带细针的注射装置向个体皮肤区域施用所述组合物所必需的性质。 As used herein, the term "injectable" refers to a material having a fine needle with an injection device using the application properties of the composition necessary to the individual area of ​​the skin. 如本文所使用,术语“细针”指27G或更小的针。 As used herein, the term "needle" refers to a 27G or smaller needle. 可通过如以上所讨论的那样确定水凝胶颗粒的尺寸实现本文公开的组合物的可注射性。 Disclosed herein may be implemented in the compositions as determined by the size of the hydrogel particles discussed above injectability.

[0089] 在该实施方案的方面,本文公开的水凝胶组合物可通过细针注射。 [0089] In aspects of this embodiment, disclosed herein, a hydrogel composition may be injected through a fine needle. 在该实施方案的其它方面,本文公开的水凝胶组合物可通过(例如)约27G、约30G或约32G的针注射。 In other aspects of this embodiment, disclosed herein, a hydrogel composition may be injected via a needle (e.g.) about a 27G, 32G to about 30G, or about. 在该实施方案的另外其它方面,本文公开的水凝胶组合物可通过(例如)22G或更小、27G或更小、 30G或更小或32G或更小的针注射。 In still other aspects of this embodiment, the herein disclosed hydrogel composition can be obtained by (e.g.) 22G or smaller, or less a 27G, 30G or 32G or smaller needle injection or smaller. 在该实施方案的另外其它方面,本文公开的水凝胶组合物可通过(例如)约22G至约35G、约22G至约34G、约22G至约33G、约22G至约32G、约22G至约27G或约27G至约32G的针注射。 In still other aspects of this embodiment, the herein disclosed hydrogel composition can be obtained by (e.g.) from about 22G to about 35G, about 22G to about 34G, about 22G to about 33G, about 22G to about 32G, about 22G to about or from about 27G injection needle 27G to about 32G of.

[0090] 在该实施方案的方面,本文公开的水凝胶组合物可用约60N、约55N、约50N、约45N、 约40N、约35N、约30N、约25N、约20N或约15N的挤压力,以100mm/min的速度注射。 [0090] In aspects of this embodiment, disclosed herein, a hydrogel composition may be about 60N, 55N about, about 50N, 45N about, about 40N, 35N about, about 30N, 25N about, about 20N to about 15N, or extrusion pressure at a speed of 100mm / min injection. 在该实施方案的其它方面,本文公开的水凝胶组合物可通过27G针,用约60N或更小、约55N或更小、约50N或更小、约45N或更小、约40N或更小、约35N或更小、约30N或更小、约25N或更小、约20N或更小、约15N或更小、约ION或更小或约5N或更小的挤压力注射。 In other aspects of this embodiment, the herein disclosed compositions may be a hydrogel 27G needle, with about 60N or less, about 55N or less, about 50N or less, about 45N or less, or about 40N , about 35N or less, about 30N or less, about 25N or less, about 20N or less, about 15N or less, about ION or less, or about 5N or less pressing force injection. 在该实施方案的另外其它方面,本文公开的水凝胶组合物可通过30G针,用约60N或更小、约55N或更小、约50N或更小、约45N或更小、约40N或更小、约35N或更小、约30N或更小、约25N或更小、约20N或更小、约15N或更小、约ION或更小或约5N或更小的挤压力注射。 In still other aspects of this embodiment, the herein disclosed compositions may be a hydrogel 30G needle, with about 60N or less, about 55N or less, about 50N or less, about 45N or less, or about 40N less, about 35N or less, about 30N or less, about 25N or less, about 20N or less, about 15N or less, about ION or less, or about 5N or less pressing force injection. 在该实施方案的另外其它方面,本文公开的水凝胶组合物可通过32G针,用约60N或更小、约55N或更小、约50N或更小、约45N或更小、 约40N或更小、约35N或更小、约30N或更小、约25N或更小、约20N或更小、约15N或更小、约ION 或更小或约5N或更小的挤压力注射。 In still other aspects of this embodiment, the herein disclosed compositions may be a hydrogel 32G needle, with about 60N or less, about 55N or less, about 50N or less, about 45N or less, or about 40N less, about 35N or less, about 30N or less, about 25N or less, about 20N or less, about 15N or less, about ION or less, or about 5N or less pressing force injection.

[0091] 本发明的方面部分地提供了本文公开的显示出粘聚性的水凝胶组合物。 [0091] aspect of the present invention provide, in part disclosed herein exhibits cohesiveness hydrogel composition. 粘聚性, 也称为内聚粘着引力、粘合力或压缩力,是材料内相似分子之间起到将分子合成一体的作用的分子间引力引起的材料的物理性质。 Cohesiveness, adhesive cohesive attraction also known as adhesive or compressive forces, similar to a play between the molecules within the material, physical properties of molecules intermolecular interaction caused integrally gravity material. 用克-力(gmf)表示粘聚性。 Represents cohesiveness force (gmf) - grams. 除其它因素外,粘结性受初始游离糖胺聚糖聚合物的分子量比、糖胺聚糖聚合物的交联度、交联后残留的游离糖胺聚糖聚合物的量和水凝胶组合物的PH影响。 Molecular weight than the degree of crosslinking of glycosaminoglycan polymer, after crosslinking of the free residual glycosaminoglycan polymer, among other factors, the initial adhesion by the amount of free polymer and glycosaminoglycan hydrogel Effect of PH compositions thereof. 组合物应有足够粘结性,以便保持位于施用部分。 The composition should have sufficient adhesion to administration holding portion located. 另外,在某些应用中,足够的粘结性对组合物保持其形状,并且因此在发生机械负载循环时保持功能性而言很重要。 Further, in some applications, sufficient adhesion is important for the composition to retain its shape, and therefore remains in terms of functionality occurs when the mechanical load cycling. 因而,在一个实施方案中,本文公开的水凝胶组合物显示出粘聚性,与水同等水平。 Thus, in one embodiment, disclosed herein hydrogel composition exhibits cohesiveness, with the same level of water. 在又一实施方案中,本文公开的凝胶组合物显示出足够粘聚性以保持位于施用部分。 In yet another embodiment, the gel compositions disclosed herein exhibit sufficient cohesiveness to maintain a portion located administered. 在又一实施方案中,本文公开的凝胶组合物显示出足够粘聚性以保持其形状。 In yet another embodiment, the gel compositions disclosed herein exhibit sufficient cohesiveness to maintain its shape. 在又一实施方案中,本文公开的凝胶组合物显示出足够粘聚性以保持其形状和功能性。 In yet another embodiment, the gel compositions disclosed herein exhibit sufficient cohesiveness to maintain its shape and functionality.

[0092] 本发明的方面部分地提供了本文公开的显示出生理上可接受的容积克分子渗透浓度(osmolarity)的水凝胶组合物。 [0092] aspect of the present invention provide, in part disclosed herein exhibit a physiologically acceptable osmolarity (osmolarity) of the hydrogel composition. 如本文所使用,术语“容积克分子渗透浓度”指溶液中渗透活性溶质的浓度。 As used herein, the term "osmolarity" refers to the activity of the solute concentration in the permeate solution. 如本文所使用,术语“生理上可接受的容积克分子渗透浓度”指与活生物体正常机能相符或其特有的容积克分子渗透浓度。 As used herein, the term "physiologically acceptable osmolarity" refers to match the osmolarity of normal functioning of living organisms or specific. 因而,在施用给哺乳动物时,施用本文公开的水凝胶组合物显示大体上没有长期或永久性有害影响的容积克分子渗透浓度。 Thus, when administered to a mammal, administering disclosed herein hydrogel composition shows no long term or permanent detrimental effect substantially osmolarity. 容积克分子渗透浓度以每升溶剂中渗透活性溶质的渗透压摩尔(Osmol/L或Osm/L)表示。 Osmolarity solvent per liter of osmotically active solute osmolarity (Osmol / L or Osm / L). FIG. 容积克分子渗透浓度与容积克分子浓度不同,因为它测量了渗透活性溶质颗粒的摩尔数而不是溶质的摩尔数。 Osmolarity and the molarity of different volume, which is measured as moles of osmotically active solute particles rather than the solute. 因为某些化合物可在溶液中离解,而其它的不能,所以出现了不同。 Because some compounds may dissociate in solution, but not the other, so there are different. 可由下列表达式计算溶液的容积克分子渗透浓度:〇ΜηοΙΖΙ^Ιφι η: C,其中Φ为渗透系数,其说明了溶液的非理想程度;η为分子离解成的颗粒(例如离子)的数量;并且c为溶质的摩尔浓度;且i为表示特定溶质同一性的指数。 Was calculated by the following expression of osmolarity: 〇ΜηοΙΖΙ ^ Ιφι η: C, wherein Φ is the permeability coefficient, which illustrates the degree of non-ideal solutions; [eta] is the number of molecules dissociated into particles (e.g., ions); and c is the molar concentration of the solute; and i is an index showing identity to a particular solute. 可使用测量溶液的常规方法测量本文公开的水凝胶组合物的容积克分子渗透浓度。 It can be measured using a conventional method of measuring a solution osmolarity disclosed herein the hydrogel composition.

[0093] 在一个实施方案中,本文公开的水凝胶组合物显示出生理上可接受的容积克分子渗透浓度。 [0093] In one embodiment, the herein disclosed hydrogel composition exhibits osmolarity physiologically acceptable. 如本文所使用,术语“重量克分子渗透浓度”指体内每千克溶剂中渗透活性溶质的浓度。 As used herein, the term "osmolality" activity refers to the concentration of solute per kilogram of solvent penetration in vivo. 如本文所使用,术语“生理上可接受的重量克分子渗透浓度”指与活生物体正常机能相符或其特有的重量克分子渗透浓度。 As used herein, the term "physiologically acceptable osmolality" refers osmolality consistent with the normal functioning of living organisms or specific. 因而,在施用给哺乳动物时,施用本文公开的水凝胶组合物显示大体上没有长期或永久性有害影响的重量克分子渗透浓度。 Thus, when administered to a mammal, administering disclosed herein hydrogel composition shows no long term or permanent detrimental effect substantially osmolality. 重量克分子渗透浓度以每千克溶剂中渗透活性溶质的渗透压摩尔(osmol/kg或Osm/kg)表示并且等于该溶液中存在的所有溶质的重量克分子浓度的总和。 Sum osmole (osmol / kg or Osm / kg) osmolality of osmotically active solute per kilogram of solvent in the solution is equal to and indicates the presence of all the molality of the solute. 可使用渗压计测量溶液的重量克分子渗透浓度。 Measured using an osmometer solution osmolality. 在现代化实验室里最常用的仪器是冰点降低渗压计。 In modern laboratories the most commonly used instrument is the freezing point depression osmometer. 该仪器测量了随重量克分子渗透浓度增加溶液中冰点发生的变化(冰点降低渗压计)或随重量克分子渗透浓度增加溶液中蒸汽压发生的变化(蒸汽压降低渗压计)。 The instrument measures the increase in osmolality with the solution freezing point variation occurring (freezing point depression osmometer) or with the change in the osmolality of the solution vapor pressure increase occurs (to reduce vapor pressure osmometer).

[0094] 在该实施方案的方面,水凝胶组合物显示出(例如)约100m0sm/L、约150m0sm/L、约200m0sm/L、约250m0sm/L、约300m0sm/L、约350m0sm/L、约400m0sm/L、约450m0sm/L 或约500m0sm/L的容积克分子渗透浓度。 [0094] In aspects of this embodiment, a hydrogel composition exhibits (e.g.) about 100m0sm / L, from about 150m0sm / L, from about 200m0sm / L, from about 250m0sm / L, from about 300m0sm / L, from about 350m0sm / L, about 400m0sm / L, from about 450m0sm / L, or from about / osmolarity of 500m0sm L. 在该实施方案的其它方面,水凝胶组合物显示出(例如) 至少100m0sm/L、至少150m0sm/L、至少200m0sm/L、至少250m0sm/L、至少300m0sm/L、至少350m0sm/L、至少400m0sm/L、至少450m0sm/L或至少500m0sm/L的容积克分子渗透浓度。 In other aspects of this embodiment, a hydrogel composition exhibits (e.g.) at least 100m0sm / least 150m0sm L, / L, at least 200m0sm / L, at least 250m0sm / L, at least 300m0sm / L, at least 350m0sm / L, at least 400m0sm / L, at least 450m0sm / L, or at least 500m0sm / L of osmolarity. 在该实施方案的另外其它方面,水凝胶组合物显示出(例如)至多100m0sm/L、至多150m0sm/L、至多200m0sm/L、至多250m0sm/L、至多300m0sm/L、至多350m0sm/L、至多400m0sm/L、至多450πι〇8Πΐ/1或至多500m0sm/L的容积克分子渗透浓度。 In still other aspects of this embodiment, a hydrogel composition exhibits (e.g.) up 100m0sm / L, up 150m0sm / L, up 200m0sm / L, up 250m0sm / L, up 300m0sm / L, up 350m0sm / L, up to L, up 450πι〇8Πΐ / or up to 1 / osmolarity 400m0sm / 500m0sm L of. 在该实施方案的另外其它方面,水凝胶组合物显不出(例如)约100m0sm/L至约500m0sm/L、约200m0sm/L至约500m0sm/L、约200m0sm/L 至约400m0sm/L、约300m0sm/L 至约400m0sm/L、约270m0sm/L 至约390m0sm/L、约225m0sm/L至约350m0sm/L、约250m0sm/L 至约325m0sm/L、约275m0sm/L至约300m0sm/L 或约285m0sm/L至约290m0sm/L的容积克分子渗透浓度。 In still other aspects of this embodiment, the hydrogel composition does not show the (e.g.) about 100m0sm / L to about 500m0sm / L, from about 200m0sm / L to about 500m0sm / L, from about 200m0sm / L to about 400m0sm / L, about 300m0sm / L to about 400m0sm / L, from about 270m0sm / L to about 390m0sm / L, from about 225m0sm / L to about 350m0sm / L, from about 250m0sm / L to about 325m0sm / L, from about 275m0sm / L to about 300m0sm / L or about 285m0sm / L to about / L osmolarity of 290m0sm.

[0095] 本发明的方面部分地提供了本文公开的显示出大体稳定性的水凝胶组合物。 Aspect of the [0095] present invention provide, in part disclosed herein exhibit substantially stable hydrogel composition. 如本文所使用,术语“稳定性”或“稳定”在提及本文公开的水凝胶组合物时,指组合物在施用给个体之前,在储存时不易于降解、分解或破碎到任何大体上或显著的程度。 As used herein, the term "stability" or "stable" in reference to the hydrogel compositions disclosed herein, refers to a composition before administration to an individual, prone to degradation during storage, disintegration or disruption of any substantially or to a significant extent. 如本文所使用, 术语“大体热稳定性”、“大体上热稳定”、“高压釜稳定”或“蒸汽灭菌稳定”指本文公开的水凝胶组合物在受到如本文公开的热处理时大体上稳定。 As used herein, the term "thermal stability generally," "substantially thermally stable", "autoclave stable" or "stable steam sterilization" means a hydrogel composition disclosed herein was subjected to heat treatment substantially as herein disclosed stable.

[0096] 可通过使水凝胶组合物受热处理,例如在正常压力或加压下(例如,高压灭菌)的蒸汽灭菌,测定本文公开的水凝胶组合物的稳定性。 [0096] can be obtained by the hydrogel composition is heat treated, for example, at normal pressure or elevated pressure (e.g., autoclaving) steam sterilization, disclosed herein determine the stability of the hydrogel composition. 优选在至少约l〇〇°C的温度下进行热处理约1分钟至约10分钟。 Heat treatment is performed preferably about 1 minute to about 10 minutes at a temperature of at least about l〇〇 ° C.. 可通过下列方式评估本文公开的水凝胶组合物的大体稳定性:1)测定灭菌后本文公开的水凝胶组合物的挤压力变化(AF),其中通过(有指定添加剂的水凝胶组合物的挤压力)减去(未加添加剂的水凝胶组合物的挤压力)测量,挤压力的变化小于2N 表明水凝胶组合物大体上稳定;和/或2)测定灭菌后本文公开的水凝胶组合物的流变性变化,其中通过(有添加剂的凝胶制剂的tanSlHz)减去(没有添加剂的凝胶制剂的tanSlHz)测量,tanSlHz的变化小于0.1表明水凝胶组合物大体上稳定。 Stability may be assessed generally disclosed herein hydrogel composition in the following ways: a pressing pressure changes) was measured after the sterilization of the hydrogel composition disclosed herein (AF), through which (with the specified additive hydrogel pressing force gum composition) is subtracted (without additives hydrogel composition pressing force) measurement, the pressing force varies by less than 2N show that the hydrogel composition is substantially stable; and / or 2) measurement changes rheology disclosed herein after sterilization hydrogel composition, wherein by (gel preparations are additive tanSlHz) subtracting (tanSlHz gel formulations without additives) measurement, tanSlHz variation of less than 0.1 indicates that a hydrogel gum composition is substantially stable. 因而,本文公开的大体上稳定的水凝胶组合物在灭菌后保持了下列一种或多种特性:均匀性、挤压力、粘结性、透明质烷浓度、试剂浓度、容积克分子渗透浓度、pH或热处理之前水凝胶所需的其它流变特性。 Accordingly, disclosed herein substantially stable hydrogel composition remains after sterilization of one or more of characteristics: uniformity, pressing force, adhesion, hyaluronan concentration, reagent concentration, molar volume osmolality, pH, or before the heat treatment required hydrogel other rheological properties.

[0097] 在一个实施方案中,使用维持本文公开的所需水凝胶性质的热处理加工包含糖胺聚糖聚合物和至少一种本文公开的试剂的水凝胶组合物。 [0097] In one embodiment, disclosed herein, used to maintain the desired heat treatment comprising a water gel properties of the hydrogel composition glycosaminoglycan polymer disclosed herein and at least one reagent. 在该实施方案的方面,使用(例如)约100 °C、约105 °C、约110 °C、约115 °C、约120 °C、约125 °C或约130 °C的热处理加工包含糖胺聚糖聚合物和至少一种本文公开的试剂的水凝胶组合物。 In aspects of this embodiment using (e.g.) heat treatment of about 100 ° C, about 105 ° C, about 110 ° C, about 115 ° C, about 120 ° C, about 125 ° C or about 130 ° C comprises a sugar hydrogel composition glycosaminoglycan polymer disclosed herein and at least one reagent. 在该实施方案的其它方面,使用(例如)至少l〇〇°C、至少105°C、至少110°C、至少115°C、至少120°C、至少125°C或至少130 °(:的热处理加工包含糖胺聚糖聚合物和至少一种本文公开的试剂的水凝胶组合物。在该实施方案的另外其它方面,使用(例如)约100°C至约120°C、约100°C至约125°C、约100°C至约130°C、约100°C至约135°C、约110°C至约120°C、约110°C至约125°C、约110°C至约130°C、约110°C 至约135°C、约120°C 至约125°C、约120°C 至约130°C、约120°C 至约135°C、约125°C 至约130°C或约125°C至约135°C的热处理加工包含糖胺聚糖聚合物和至少一种本文公开的试剂的水凝胶组合物。 In other aspects of this embodiment using (e.g.) at least l〇〇 ° C, at least 105 ° C, at least 110 ° C, at least 115 ° C, at least 120 ° C, at least 125 ° C or at least 130 ° (: the heat treatment comprising glycosaminoglycan polymer disclosed herein and at least one agent hydrogel composition. in yet other aspects, this embodiment (e.g.) about between 100 ° C to about 120 ° C, from about 100 ° C to about 125 ° C, approximately between 100 ° C to about 130 ° C, approximately between 100 ° C to about 135 ° C, about 110 ° C to about 120 ° C, about 110 ° C to about 125 ° C, about 110 ° C to about 130 ° C, about 110 ° C to about 135 ° C, about 120 ° C to about 125 ° C, about 120 ° C to about 130 ° C, about 120 ° C to about 135 ° C, about 125 ° C to about 130 ° C or about 125 ° C to 135 ° C to about heat treatment comprising a glycosaminoglycan polymer disclosed herein and at least one agent hydrogel composition.

[0098] 可通过使水凝胶组合物受热处理,例如储存在约45 °C环境中约60天,测定本文公开的水凝胶组合物的长期稳定性。 [0098] can be obtained by the hydrogel composition by heat treatment, such as storage at about 45 ° C environment for about 60 days, and the long term stability disclosed herein, the hydrogel composition. 可通过下列方式评估本文公开的水凝胶组合物的长期稳定性:1)估计45°C热处理后水凝胶组合物的清澈度和颜色,水凝胶组合物清澈且未着色表明水凝胶组合物大体上稳定;2)测定在45°C热处理后本文公开的水凝胶组合物的挤压力变化(AF),其中通过(45°C热处理之前,有指定添加剂的水凝胶组合物的挤压力)减去(45°C 热处理之后,有指定添加剂的水凝胶组合物的挤压力)测量,挤压力的变化小于2N表明水凝胶组合物大体上稳定;和/或3)测定灭菌后本文公开的水凝胶组合物的流变性变化,其中通过(45°C热处理之前,有指定添加剂的凝胶制剂的tanSlHz)减去(45°C热处理之后,有指定添加剂的凝胶制剂的tan5IHz)测量,tan5IHz的变化小于0.1表明水凝胶组合物大体上稳定。 Disclosed herein may assess long-term stability of the hydrogel composition in the following ways: 1) estimation clarity and color after heat treatment 45 ° C aqueous gel compositions, hydrogel compositions show clear and not colored hydrogel the composition is substantially stable; 2) measuring a change in pressing force after 45 ° C heat treatment disclosed herein, a hydrogel composition (the AF), which by prior (45 ° C heat treatment, the hydrogel composition designated additives after pressing force) minus (45 ° C heat treatment, there is a pressing force hydrogel composition designated additive) measurement, the pressing force varies by less than 2N show that the hydrogel composition is substantially stable; and / or 3) after the change in rheological herein disclosed hydrogel composition was measured after sterilization, which by prior (45 ° C heat treatment, the gel formulation has tanSlHz specified additive) subtracting (45 ° C heat treatment, the additive has designated the gel formulation tan5IHz) measuring the change tan5IHz less than 0.1 indicates that the hydrogel composition is substantially stable. 因而,用在45°C热处理之后保持了下列一种或多种特征评估本文公开的水凝胶组合物的长期稳定性:清澈度(透明和半透明)、均匀性和粘结性。 Thus, the holding of one or more of the following characteristics after heat treatment is 45 ° C assess long-term stability of the herein disclosed hydrogel composition: clarity (transparent and translucent), uniformity and adhesion.

[0099] 在该实施方案的方面,水凝胶组合物在室温下大体上稳定(例如)约3个月、约6个月、约9个月、约12个月、约15个月、约18个月、约21个月、约24个月、约27个月、约30个月、约33个月或约36个月。 [0099] In aspects of this embodiment, a hydrogel composition is substantially stable (e.g.) about 3 months, about 6 months, about 9 months, about 12 months, about 15 months, at about room temperature 18 months, about 21 months, about 24 months, about 27 months, about 30 months, about 33 months, or about 36 months. 在该实施方案的其它方面,水凝胶组合物在室温下大体上稳定(例如) 至少3个月、至少6个月、至少9个月、至少12个月、至少15个月、至少18个月、至少21个月、至少24个月、至少27个月、至少30个月、至少33个月或至少36个月。 In other aspects of this embodiment, a hydrogel composition is substantially stable at room temperature (e.g.) at least 3 months, at least 6 months, at least 9 months, at least 12 months, at least 15 months, at least 18 months, at least 21 months, at least 24 months, at least 27 months, at least 30 months, at least 33 months, or at least 36 months. 在该实施方案的其它方面, 水凝胶组合物在室温下大体上稳定(例如)约3个月至约12个月、约3个月至约18个月、约3个月至约24个月、约3个月至约30个月、约3个月至约36个月、约6个月至约12个月、约6个月至约18个月、约6个月至约24个月、约6个月至约30个月、约6个月至约36个月、约9个月至约12 个月、约9个月至约18个月、约9个月至约24个月、约9个月至约30个月、约9个月至约36个月、 约12个月至约18个月、约12个月至约24个月、约12个月至约30个月、约12个月至约36个月、 约18个月至约24个月、约18个月至约30个月或约18个月至约36个月。 In other aspects of this embodiment, a hydrogel composition is substantially stable at room temperature (e.g.) about 3 months to about 12 months, about 3 months to about 18 months, about 3 months to about 24 months, about 3 months to about 30 months, about 3 months to about 36 months, about 6 months to about 12 months, about 6 months to about 18 months, about 6 months to about 24 months, about 6 months to about 30 months, about 6 months to about 36 months, about 9 months to about 12 months, about 9 months to about 18 months, about 9 months to about 24 months, about 9 months to about 30 months, about 9 months to about 36 months, about 12 months to about 18 months, about 12 months to about 24 months, about 12 months to about 30 months, about 12 months to about 36 months, about 18 months to about 24 months, about 18 months to about 30 months or about 18 months to about 36 months.

[0100] 本组合物可任选包括但不限于其它药学上可接受的组分,包括但不限于缓冲剂、 防腐剂、张力调节剂、盐、抗氧化剂、重量克分子渗透浓度调节剂、乳化剂、湿润剂等。 [0100] The present compositions may optionally include but are not limited to, other pharmaceutically acceptable components, including, but not limited to, buffers, preservatives, tonicity adjusting agents, salts, antioxidants, osmolality adjusting agents, emulsifying agents, wetting agents and the like.

[0101] 药学上可接受的缓冲剂是可用于制备本文公开的水凝胶组合物的缓冲剂,条件是所得制剂在药学上可接受。 [0101] Pharmaceutically acceptable buffers may be used for preparing buffer is herein disclosed hydrogel composition, with the proviso that the resulting in a pharmaceutically acceptable formulation. 药学上可接受的缓冲剂的非限制性实例包括乙酸盐缓冲剂、硼酸盐缓冲剂、柠檬酸盐缓冲剂、中性缓冲盐水、磷酸盐缓冲剂和磷酸盐缓冲盐水。 Pharmaceutically acceptable non-limiting examples of buffers include acetate buffers, borate buffers, citrate buffers, neutral buffered saline, phosphate buffered saline, and phosphate buffers. 可将任何浓度的药学上可接受的缓冲剂用于配制本文公开的药物组合物,条件是使用有效浓度的缓冲剂恢复治疗有效量的活性成分。 Any pharmaceutically acceptable buffer concentration can be used to formulate the pharmaceutical compositions disclosed herein, with the proviso that the use of effective concentrations of buffering agents to restore a therapeutically effective amount of the active ingredient. 生理上可接受的缓冲剂浓度的非限制性实例存在于约0. ImM至约900mM范围内。 Physiologically acceptable non-limiting examples of buffering agents is present in a concentration from about 0. ImM to about 900mM range. 可调节药学上可接受的缓冲剂的pH,条件是所得制剂是在药学上可接受。 Adjustable pharmaceutically acceptable buffer the pH, provided that the resulting preparation is pharmaceutically acceptable on. 应理解可根据需要使用酸或碱调节药物组合物的pH。 It should be understood that acids or bases may be used adjust the pH of the pharmaceutical composition required. 可将任何缓冲的pH水平用于配制药物组合物,条件是使用该有效PH水平恢复治疗有效量的母体聚合物。 Any buffer pH levels can be formulated to pharmaceutical compositions, that use of the effective horizontal recovery PH therapeutically effective amount of matrix polymer. 生理上可接受的pH的非限制性实例存在于约pH 5.0至约pH 8.5的范围内。 Physiologically acceptable non-limiting examples of pH present in the range of 5.0 to about pH 8.5 to about pH. 例如,本文公开的水凝胶组合物的pH可为约5.0至约8.0或约6.5至约7.5、约7.0至约7.4或约7.1至约7.3。 For example, pH of the hydrogel composition disclosed herein may be from about 5.0 to about 8.0, or from about 6.5 to about 7.5, from about 7.0 to about 7.4, or from about 7.1 to about 7.3.

[0102] 药学上可接受的防腐剂包括但不限于焦亚硫酸钠、硫代硫酸钠、乙酰半胱氨酸、丁羟茴香醚和丁羟甲苯。 [0102] Pharmaceutically acceptable preservatives include, but are not limited to, sodium metabisulfite, sodium thiosulfate, acetylcysteine, butylated hydroxyanisole and butylated hydroxytoluene. 药学上可接受的防腐剂包括但不限于苯扎氯铵、氯丁醇、硫柳汞、醋酸苯萊、硝酸苯萊、稳定化氧氯组合物,例如PlJRFTEi' (Allergan,Inc. Irvine,CA)和螯合剂,例如DTPA或DTPA-双酰胺、DTPA钙及CaNaDTPA-双酰胺。 Pharmaceutically acceptable preservatives include, but are not limited to, benzalkonium chloride, chlorobutanol, thimerosal, phenylmercuric acetate, phenylmercuric Levin, Levin, phenylmercuric nitrate, a stabilized oxychloro composition, e.g. PlJRFTEi '(Allergan, Inc. Irvine, CA), and chelating agents such as DTPA or DTPA- bisamide, calcium DTPA-bisamide and CaNaDTPA-.

[0103] 用于本文公开的水凝胶组合物中的药学上可接受的张力调节剂包括但不限于盐例如氯化钠和氯化钾;和甘油。 [0103] Pharmaceutically used herein disclosed hydrogel composition in a pharmaceutically acceptable tonicity adjusting agents include, but are not limited to, salts such as sodium chloride and potassium chloride; and glycerol. 可将所述组合物作为盐提供并且可与许多酸一起形成,包括但不限于,盐酸、硫酸、乙酸、乳酸、酒石酸、苹果酸、琥珀酸等。 The composition may be provided as a salt and can be formed with many acids, including but not limited to, hydrochloric, sulfuric, acetic, lactic, tartaric, malic, succinic, etc. 盐倾向于比相应的游离碱形式更易溶于水或其它质子溶剂。 Salts tend to be more soluble in water or other protic solvents than the corresponding free base forms. 应理解,在本文公开的药物组合物中可包括药学领域内已知的这些和其它物质。 It should be understood, herein disclosed pharmaceutical composition may comprise these and other substances known in the art of pharmacy. 药学上可接受的组分的其它非限制性实例可在例如Ansel,同上, (1999) ;Gennaro,同上,(2000) ;Hardman,同上,(2001);和Rowe,同上,(2003)中找到,由此将其中每一个通过引用的方式整体并入。 Other non-limiting examples of pharmaceutically acceptable components may be, for example, Ansel, supra, (1999); Gennaro, supra, (2000); Hardman, supra, (2001); and Rowe, supra, (2003) found , whereby each of which is incorporated by reference in its entirety.

[0104] 本发明的方面部分地提供了通过施用本文公开的水凝胶组合物治疗个体的软组织状况的方法。 [0104] aspect of the present invention provides, in part by administering a hydrogel composition disclosed herein the treatment of soft tissue in the condition of the individual. 如本文所使用,术语“治疗”指减轻或消除个体中特征在于软组织缺陷、缺损、疾病和/或病症的软组织状况的美容或临床症状;或延迟或预防个体中特征在于软组织缺陷、缺损、疾病和/或病症的状况的美容或临床症状发作。 As used herein, the term "treatment" refers to reducing or eliminating the individual characteristics in that a cosmetic or clinical symptoms of soft tissue condition of soft tissue defects, defect, disease and / or condition; or delaying or preventing the characteristics of the individual wherein the soft tissue defect, defect, disease status and / or condition of beauty or onset of clinical symptoms. 例如,术语“治疗”可指减轻特征在于软组织缺损、疾病和/或病症的状况的症状,例如至少20%、至少30%、至少40%、至少50%、至少60%、至少70%、至少80%、至少90%或至少100%。 For example, the term "treating" may refer to reduce wherein soft tissue defects, symptoms and condition and / or disorder diseases, such as at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90% or at least 100%. 可通过观测一种或多种美容、 临床症状和/或与所述状况相关的生理指标测定本文公开的水凝胶组合物在治疗特征在于软组织缺损、疾病和/或病症的状况中的效果。 By observing one or more cosmetic, clinical symptoms and / or condition related to the measured physiological parameters disclosed herein, wherein a hydrogel composition of soft tissue defects, the effect of the disease state and / or in the treatment of disorders of the features. 也可由对同步疗法的需要减少表明软组织缺损、疾病和/或病症改善。 But also by reducing the need for synchronization therapy showed soft tissue defects, diseases and / or conditions improve. 本领域的技术人员将了解与特定软组织缺损、疾病和/或病症相关的适当症状或指标并且将了解如何确定个体是否是用本文公开的化合物或组合物治疗的候选人。 Those skilled in the art will appreciate that the specific soft tissue defect, or a suitable indicator of disease symptoms and / or disorders associated and will know how to determine a candidate therapeutic compound or composition of whether an individual is disclosed herein.

[0105] 向个体施用根据本发明所述的水凝胶组合物。 [0105] The hydrogel composition is administered according to the invention to a subject. 个体通常是任何年龄、性别或种族的人类。 Individuals often a human of any age, sex or race. 通常,是常规程序治疗软组织状况的候选人的任何个体也是本文公开的方法的候选人。 Typically, a candidate of any method disclosed herein is an individual program routine treatment of soft tissue conditions of the candidates. 虽然体验到皮肤老化征兆的受试者是成人,但是体验到适合治疗的过早老化或其它皮肤状况(例如,疤痕)的受试者也可用本文公开的水凝胶组合物治疗。 Although signs of aging of the skin to experience the subject is an adult, but experience suitable for the treatment of premature aging or other skin conditions (e.g., scarring) may also be subject therapeutic hydrogel composition disclosed herein. 另外,目前公开的水凝胶组合物和方法可应用于寻求身体部位或区域的小/中度膨大、形状变化或轮廓变更的个体,这用现有软组织植入技术,在技术上不可能或在审美上不能接受。 Moreover, hydrogel compositions and methods of the presently disclosed may be applied to a body part or region to seek small / moderate swelling, changes in the shape or contour of the individual change, with this prior art implant soft tissue, technically impossible or unacceptable aesthetically. 除全面告知应允公开所述程序的所有相关风险和利益外,术前评估通常包括常规史和身体检查。 In addition to a comprehensive program to inform the public answered all relevant risks and benefits of routine preoperative evaluation usually includes a history and physical examination.

[0106] 本文公开的水凝胶组合物和方法对治疗软组织状况有用。 [0106] The hydrogel compositions and methods disclosed herein are useful for the treatment of soft tissue conditions. 软组织状况包括但不限于软组织缺陷、缺损、疾病和/或病症。 Soft tissue condition including, but not limited to soft tissue defects, defects, diseases and / or disorders. 软组织状况的非限制性实例包括乳房缺陷、缺损、疾病和/或病症,例如乳房增大、乳房再造、乳房固定、乳房过小、胸部发育不全、波兰氏综合征(Poland's syndrome)、由于植入并发症引起的缺损如囊袋收缩和/或破裂;面部缺陷、缺损、疾病或病症,例如面部增大、面部再造、美索疗法、帕-罗二氏综合征(Parry-Romberg syndrome)、深部红斑狼疮、真皮麻点、疤痕、面颊凹陷、薄唇、鼻部缺陷或缺损、眶后缺陷或缺损、面部褶皱、细纹和/或皱纹,如眉间纹、鼻唇纹、口周纹和/或嘴角纹,和/或面部的其它轮廓畸形或缺陷;颈部缺陷、缺损、疾病或/或病症;皮肤缺陷、缺损、疾病或/或病症;其它软组织缺陷、缺损、疾病或/或病症,例如上臂、下臂、手、肩膀、背部、躯干(包括腹部)、臀部、大腿、小腿(包括腓)、脚(包括足底脂肪垫)、眼睛、生殖器或其它身体部 Non-limiting examples of conditions include breast soft tissue defects, defects, diseases and / or disorders, e.g. breast augmentation, breast reconstruction, mastopexy, breast is too small, breast hypoplasia, Poland's syndrome (Poland's syndrome), since the implant complications such as defects caused by bladder contractions and / or rupture; facial defects, defect, disease or disorder, such as the face increases, facial reconstruction, Mae Sot therapy, Pa - Luoer Shi syndrome (Parry-Romberg syndrome), deep lupus erythematosus, leather pitting scars, cheek depressions, thin lips, nasal defect or defects, flaws or defects orbital facial wrinkles, fine lines and / or wrinkles, such as glabellar lines, nasolabial folds, perioral lines and / or marionette lines, and / or other facial contour deformities or defect; neck defects, defect, disease or / or disorders; skin imperfections, defect, disease or / or condition; other soft tissue defects, defect, disease or / or disorders such as upper arms, lower arms, hands, shoulders, back, torso (including the abdomen), buttocks, thighs, legs (including Phil), feet (including plantar fat pad), eyes, genitals or other body part 、区域或面积的增大或再造,或影响这些身体部位、区域或面积的疾病或病症;小便失禁、大便失禁、其它形式的失禁;和胃食管反流疾病(GERD)。 , Region or area augmentation or reconstruction, or the impact of these body parts, the region or area of ​​the disease or disorder; urinary incontinence, fecal incontinence, other forms of incontinence; and gastroesophageal reflux disease (GERD). 如本文所使用,术语“美索疗法”指皮肤的非手术美容治疗技术,牵涉向表皮、真皮-表皮交界处和/或真皮经表皮内、真皮内和/或皮下注射呈多个小液滴施用的试剂。 As used herein, the term "Mae Sot therapy" refers to a non-surgical cosmetic skin treatment techniques, involving the epidermis, dermal - epidermal junction and / or dermis via intraepidermal, intradermal and / or subcutaneous injection as a plurality of small droplets application of reagents.

[0107] 通常将基于所需改变和/或改善、所需软组织状况症状的减轻和/或消除、个体和/ 或医师所需临床和/或美容效果和治疗的身体部位或区域确定与本文公开的任何方法一起使用的水凝胶组合物的量。 [0107] typically based on the desired changes and / or improvements to alleviate the symptoms of the condition required for soft tissue and / or eliminate the required individual and / or physicians in clinical and / or body part or area of ​​cosmetic results and determine treatment disclosed herein any amount of aqueous gel composition for use with the methods. 组合物施用的效果可用下列一种或多种临床和/或美容度量标准表示:软组织形状改变和/或改善、软组织尺寸改变和/或改善、软组织轮廓改变和/或改善、组织功能改变和/或改善、组织向内生长支持和/或新胶原蛋白沉积、组合物持续灌输、 患者满意度和/或生活质量提高及可植入异物的使用减少。 The composition administered effect can be used one or more of clinical and / or cosmetic metric represents: soft tissue shape change and / or improve the soft tissue size change and / or improve the soft tissue profile change and / or improve tissue function changes and / or amelioration supports tissue ingrowth and / or new collagen deposition, continuous infusion composition, patient satisfaction and / or quality of life and reduce the use of implantable foreign matter.

[0108] 所述组合物和方法治疗面部软组织的效果可用下列一种或多种临床和/或美容度量标准表示:面部特征的尺寸、形状和/或轮廓改善,如嘴唇、面颊或眼部区域的尺寸、形状和/或轮廓改善;面部特征的尺寸、形状和/或轮廓改变,如嘴唇、面颊或眼部区域的尺寸、形状和/或轮廓改变;皮肤上的皱纹、褶皱或细纹减少或消除;对皮肤上的皱纹、褶皱或细纹有抗性;皮肤补水;皮肤弹性增加;皮肤粗糙减轻或消除;皮肤紧固度增加和/或改善;拉伸纹或妊娠纹减少或消除;肤色、光泽、亮度和/或光彩增强和/或改善;皮肤颜色增强和/或改善、皮肤苍白减轻或消除;组合物持续灌输;副作用减少;患者满意度和/或生活质量提高。 [0108] The effects of compositions and methods for treating soft tissue of the face can be used one or more of clinical and / or cosmetic metric represents: facial feature size, shape, and / or to improve the profile, such as the lips, cheek or eye region the size, shape and / or profile improving; facial feature size, shape and / or contour changes, such as the size of the lips, cheek or eye region, the shape and / or profile change; wrinkles, fine lines or wrinkles on the skin to reduce or elimination; resistant to wrinkles, fine lines or wrinkles on the skin; skin replenishment; increase in skin elasticity; reduce or eliminate rough skin; skin tautness increased and / or improved; stretch marks, or reduction or elimination of stretch marks; color, gloss, brightness and / or luster enhance and / or improve; skin color enhancement and / or improve, reduce or eliminate pale skin; composition continued to instill; reduction of side effects; patient satisfaction and / or quality of life.

[0109] 作为又一个例子,对于小便失禁手术而言,用于括约肌支持的组合物和方法的效果可用下列一种或多种临床度量标准表示:失禁频率减小、持续灌输、患者满意度和/或生活质量提高及可植入外来填充剂的使用减少。 [0109] As a further example, for urinary incontinence surgery, the effect of the composition and methods for the sphincter to support one or more of the available clinical metric represents: incontinence frequency decreases, continuous infusion, patient satisfaction and / or improve the quality of life and the use of implantable foreign filler reduced.

[0110] 在该实施方案的方面,施用的水凝胶组合物的量为(例如)约〇. Olg、约〇. 〇5g、约0. lg、约0.5g、约lg、约5g、约10g、约20g、约30g、约40g、约50g、约60g、约70g、约80g、约90g、 约100g、约150g或约200g。 [0110] In aspects of this embodiment, the amount administered hydrogel composition (e.g.) about billion. OLG, approximately square. 〇5g, about of 0. The lg, about 0.5g, about lg, about 5g, about 10g, about 20g, about 30g, about 40g, about 50g, about 60g, about 70g, about 80g, about 90g, about 100g, or from about 150g to about 200g. 在该实施方案的其它方面,施用的水凝胶组合物的量为(例如)约0 .Olg至约0. lg、约0. Ig至约lg、约Ig至约10g、约IOg至约IOOg或约50g至约200g。 In other aspects of this embodiment, the amount administered hydrogel composition (e.g.) from about to about of 0. The 0 .Olg lg, from about Ig to about of 0. The lg, from about Ig to about 10g, about IOg about IOOg or about 50g to about 200g. 在该实施方案的另外其它方面,施用的水凝胶组合物的量为(例如)约〇. 〇ImL、约0.05mL、约0. ImL、约0.5mL、约lmL、约5mL、约10mL、约20mL、约30mL、约40mL、约50mL、约60mL、约70g、约80mL、约90mL、约I OOmL、约150mL或约200mL。 In still other aspects of this embodiment, the amount administered hydrogel composition (e.g.) about billion. 〇ImL, about 0.05mL, about 0. ImL, about 0.5 mL, about lmL, about 5mL, about 10mL, about 20mL, about 30mL, about 40mL, about 50mL, about 60mL, about 70g, about 80mL, about 90mL, about I OOmL, about 150mL, or about 200mL. 在该实施方案的其它方面,施用的水凝胶组合物的量为(例如)约〇· 〇I mL至约0 · I mL、约0 · I mL至约I mL、约I mL至约I OmL、约I OmL至约10 OmL或约5 OmL 至约200mL。 In other aspects of this embodiment, the amount administered hydrogel composition (e.g.) about billion-〇I mL to about I 0-mL, from about 0 mL to about I-I mL, from about I to about I mL OmL, from about to about 10 OmL I OmL, or about 5 OmL to about 200mL.

[0111] 通常将基于个体和/或医师所需的美容和/或临床效果及治疗的身体部位或区域测定治疗的持续时间。 [0111] The duration of the treatment is generally based on individual measurement and / or the desired cosmetic physician and / or body part or area of ​​treatment and clinical effect. 在该实施方案的方面,施用本文公开的水凝胶组合物可治疗软组织状况,例如约6个月、约7个月、约8个月、约9个月、约10个月、约11个月、约12个月、约13个月、 约14个月、约15个月、约18个月或约24个月。 In aspects of this embodiment, the administering disclosed herein hydrogel composition to treat soft tissue condition, for example, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, about 12 months, about 13 months, about 14 months, about 15 months, about 18 months or about 24 months. 在该实施方案的其它方面,施用本文公开的水凝胶组合物可治疗软组织状况,例如至少6个月、至少7个月、至少8个月、至少9个月、至少10 个月、至少11个月、至少12个月、至少13个月、至少14个月、至少15个月、至少18个月或至少24个月。 In other aspects of this embodiment, the administering disclosed herein hydrogel composition to treat soft tissue condition, such as at least 6 months, at least seven months, at least eight months, at least 9 months, at least 10 months, at least 11 months, at least 12 months, at least 13 months, at least 14 months, at least 15 months, at least 18 months, or at least 24 months. 在该实施方案的其它方面,施用本文公开的水凝胶组合物可治疗软组织状况,例如约6个月至约12个月、约6个月至约15个月、约6个月至约18个月、约6个月至约21个月、约6个月至约24个月、约9个月至约12个月、约9个月至约15个月、约9个月至约18个月、约9个月至约21个月、约6个月至约24个月、约12个月至约15个月、约12个月至约18个月、约12个月至约21个月、约12个月至约24个月、约15个月至约18个月、约15个月至约21个月、约15个月至约24个月、约18个月至约21个月、约18个月至约24个月或约21个月至约24个月。 In other aspects of this embodiment, a hydrogel composition is administered disclosed herein to treat soft tissue condition, for example from about 6 months to about 12 months, about 6 months to about 15 months, about 6 months to about 18 months, about 6 months to about 21 months, about 6 months to about 24 months, about 9 months to about 12 months, about 9 months to about 15 months, about 9 months to about 18 months, about 9 months to about 21 months, about 6 months to about 24 months, about 12 months to about 15 months, about 12 months to about 18 months, about 12 months to about 21 months, about 12 months to about 24 months, about 15 months to about 18 months, about 15 months to about 21 months, about 15 months to about 24 months, about 18 months to about 21 months, about 18 months to about 24 months or about 21 months to about 24 months.

[0112] 本发明的方面部分地提供了施用本文公开的水凝胶组合物。 Aspect of the [0112] present invention provide, in part disclosed herein administering a hydrogel composition. 如本文所使用,术语“施用”指向个体提供本文公开的组合物的任何递送机制,其潜在地导致了临床上、在治疗上或实验上有益的结果。 As used herein, the term "administration" to an individual to provide any delivery mechanism disclosed herein is a composition, which potentially leads to clinical benefit in the treatment or experimental results. 用于向个体施用组合物的实际递送机制可通过本领域普通技术人员考虑包括但不限于以下的因素来确定:皮肤状况的类型、皮肤状况的位置、皮肤状况的原因、皮肤状况的严重性、所需缓解程度、所需缓解的持续时间、使用的特定组合物、使用的特定组合物的排泄率、使用的特定组合物的药效、使用的特定组合物中包括的其它化合物的性质、特定施用途径、特定特性、个体病史和风险因素(例如年龄、体重、健康状况等)或其任何组合。 The actual delivery mechanism for administering the composition may include, but are not limited to be determined by factors of ordinary skill in the art in view: Severity reason type of skin condition, the position of skin conditions, skin conditions, skin conditions, extent alleviate the desired properties, the duration of relief desired, the particular composition, the rate of excretion of the particular composition used, the efficacy of the particular composition used, and other compounds used in the particular composition comprising the specific route of administration, the particular characteristics, history and individual risk factors (such as age, weight, health, etc.) or any combination thereof. 在该实施方案的一个方面,通过注射向个体的皮肤区域施用本文公开的组合物。 In one aspect of this embodiment, the composition disclosed herein is administered to the skin region of a subject by injection.

[0113] 通常将基于个体和/或医师所需美容和/或临床效果和治疗的身体部位或区域确定向个体患者施用水凝胶组合物的途径。 Body part or area [0113] Generally the desired cosmetic and / or clinical effects and treatment based on individual and / or physician determines the route hydrogel composition is administered to the individual patient. 本文公开的组合物可通过本领域普通技术人员已知的任何方式施用,包括但不限于带针注射器、手枪(例如,液压气动压缩手枪)、导管、局部或通过直接手术植入。 Any embodiment disclosed herein may be a composition known to those of ordinary skill in the art by administration, including but not limited to a syringe with a needle, pistol (e.g., compression hydropneumatic gun), a catheter, or by direct surgical implantation locally. 可向皮肤区域,例如真皮区或皮下区施用本文公开的水凝胶组合物。 Area of ​​the skin may be, for example, the dermal or subcutaneous administration region hydrogel composition disclosed herein. 例如,可利用直径约0.26mm至约0.4mm且长度范围从约4mm至约14mm的针注射本文公开的水凝胶组合物。 For example, by using a diameter of about 0.26mm to about 0.4mm and a length ranging from about 4mm to about 14mm of needle injection herein disclosed hydrogel composition. 可选地,针可为21-32G并且长度为约4mm至约70mm。 Alternatively, the needle may be 21-32G and a length of about 4mm to about 70mm. 优选地,针为一次性使用针。 Preferably, the needle is a disposable needle. 所述针可与注射器、导管和/或手枪组合。 The needle, a catheter, and / or in combination with a syringe pistol.

[0114] 另外,本文公开的组合物可一次或分多次施用。 [0114] Further, herein disclosed compositions may be administered once or more times. 最后,使用的定时将按照护理质量标准。 Finally, the timing will be used in accordance with the quality of care standards. 例如,本文公开的水凝胶组合物可一次或分几个时段施用,时段间隔几天或几周。 For example, disclosed herein, a hydrogel composition may be administered once or divided into several periods, the period interval of days or weeks. 例如,个体可每1、2、3、4、5、6或7天或每1、2、3或4周施用本文公开的水凝胶组合物。 For example, an individual may be administered or hydrogel composition disclosed herein 1,2,3,4,5,6, or 7 days per every 3 or 4 weeks. 向个体施用本文公开的水凝胶组合物可一个月或两个月一次或每3、6、9或12个月施用一次。 Disclosed herein is administered to an individual hydrogel composition may be one or two months, or every 3,6, 9, or 12 months, administered once again.

[0115] 本发明的方面部分地提供了真皮区。 Aspect of the [0115] present invention provide, in part dermal region. 如本文所使用,术语“真皮区”指包含表皮-真皮交界处的皮肤和包括浅表真皮(乳头区)和深层真皮(网状区)的真皮的区域。 As used herein, the term "dermal region" refers to an epidermal - dermal junction region and comprising a superficial dermis (papillary region) and the deep dermis (reticular region) dermis. 皮肤由三个主要的层组成:表皮,其提供了防水性并用作感染的屏障;真皮,其用作皮肤附属物的位置; 和皮下组织(皮下脂肪层)。 The skin consists of three main layers: the epidermis, which provides waterproofing and serves as a barrier to infection; the dermis, which serves as the position of the skin appendages; and subcutis (subcutaneous fat layer). 表皮不含血管,并通过从真皮的扩散得到滋养。 Avascular epidermis, obtained by diffusion from the dermis and nourishment. 组成表皮的主要细胞类型为角化细胞、黑素细胞、朗格汉斯细胞(Langerhans cell)和梅克尔细胞(Merkels cell) 〇 The main cell types of epidermal keratinocytes, melanocytes, Langerhans cells (Langerhans cell) and Merkel cell (Merkels cell) square

[0116] 真皮是表皮下由结缔组织组成的皮肤层并为身体缓冲应力和应变。 [0116] The dermis is the layer of skin beneath the epidermis, and a buffer composed of connective tissue stress and strain for the body. 真皮与表皮通过基膜紧紧连接。 Dermis and epidermis firmly connected by a basement membrane. 它也包含提供触感和热感的许多机械性刺激感受器/神经末梢。 It also contains a number of mechanical and thermal provide tactile stimulation of receptor / nerve endings. 它含有毛囊、汗腺、皮脂腺、顶泌腺、淋巴管和血管。 It contains hair follicles, sweat glands, sebaceous glands, apocrine glands, lymphatic vessels and blood vessels. 在真皮中的血管提供了营养并且从它自身的细胞以及从表皮的基底层去除废物。 Blood vessels in the dermis provides nutrients and removes wastes from the basal layer of the epidermis as well as from its own cells. 真皮在结构上分为两个区:邻近表皮、称为乳头区的浅表区和称为网状区的深部较厚区。 Dermis is structurally divided into two areas: adjacent to the epidermis, called the papillary region, and superficial zone deep thicker area known as the reticular region.

[0117] 乳头区由松弛网形结缔组织构成。 [0117] nipple-shaped region consists of connective tissue web slack. 它因其指状突出物而被称为乳头,其向表皮延伸。 It is because the finger is referred to as a nipple projection which extends epidermis. 乳头为真皮提供了与表皮互相交错的“崎岖不平的”表面,加强了两层皮肤之间的连接。 Papillary dermis provides the epidermis with mutually interleaved "bumpy" surface, to strengthen the connection between the two skins. 网状区位于乳头区深处,并且通常厚很多。 Mesh area is located deep in the nipple area, and usually much thicker. 它由致密的不规则结缔组织构成,并由编织它的致密浓度的胶原、弹性和网状纤维得名。 It is composed of dense irregular connective tissue, by which a dense weave concentration collagen, elastic fibers and reticular name. 这些蛋白纤维给予真皮强度、延展性和弹性性质。 These proteins leather fibers give strength, ductility and elastic properties. 也位于网状区之内的是毛发根部、皮脂腺、汗腺、感受器、指甲和血管。 Also within the mesh zone are the roots of the hair, sebaceous glands, sweat glands, receptors, nails and blood vessels. 纹身油墨保留在真皮中。 Tattoo ink is retained in the dermis. 由怀孕产生的妊娠纹也位于真皮中。 Stretch marks from pregnancy, also located in the dermis.

[0118] 皮下组织位于真皮下面。 [0118] located in the subcutaneous tissue beneath the dermis. 其作用是连接皮肤真皮区与下面的骨和肌肉以及为它提供血管和神经。 Its role is to connect the dermis of the skin area and underlying bone and muscle as well as blood vessels and nerves to provide it. 它由疏松结缔组织和弹性蛋白组成。 It consists of loose connective tissue and elastin. 主要的细胞类型为成纤维细胞、巨噬细胞和脂细胞(皮下组织含有50%的体脂)。 The main cell types of fibroblasts, macrophages and adipocytes (hypodermis contains 50% body fat). 脂肪用作身体的垫料(padding)和保温材料。 Body fat as litter (padding), and insulation materials.

[0119] 在该实施方案的一个方面,通过向皮下区域的真皮区注射,向个体的皮肤区域施用本文公开的水凝胶组合物。 [0119] In one aspect of this embodiment, by injection into the subcutaneous region of the dermal region, administration of the hydrogel compositions disclosed herein to an area of ​​skin of an individual. 在该实施方案的方面,通过向(例如)表皮-真皮交界区、乳头区、网状区或其任何组合注射,向个体的真皮区施用本文公开的水凝胶组合物。 In aspects of this embodiment, by the (e.g.) epidermis - disclosed dermal junction region, the nipple area, region, or any combination thereof reticular injection, dermal administration to an individual region hydrogel composition described herein.

[0120] 有利的是,本发明一些组合物对减少(例如)患者薄皮区内细纹的出现特别有用且有效。 [0120] Advantageously, some compositions of the present invention (e.g.) lines occurs in patients with thin skin region is particularly useful and effective in reducing. 例如,提供了细纹治疗的方法,包括在不大于约Imm的深度,向患者施用本文其它地方描述的真皮填充剂组合物的步骤。 For example, a method of treating fine lines, including a depth of no more than about Imm, step dermal filler composition as described elsewhere herein is administered to a patient.

[0121] 本说明书的其它方面部分地公开了治疗皮肤状况的方法,包括向患有皮肤状况的个体施用本文公开的水凝胶组合物的步骤,其中施用所述组合物改善了皮肤状况,从而治疗了皮肤状况。 [0121] Additional aspects of the present specification discloses a method of treating a skin condition, comprising the steps herein disclosed hydrogel composition is administered to an individual suffering from a skin condition, wherein the administration of the composition improves skin condition, thereby treatment of the skin condition. 在该实施方案的一个方面,(皮肤状况是)治疗皮肤脱水的方法包括向患有皮肤脱水的个体施用本文公开的水凝胶组合物的步骤,其中施用所述组合物为皮肤补水, 从而治疗了皮肤脱水。 In one aspect of this embodiment, (a skin condition) The method of treating skin comprising the step of dehydration disclosed herein the hydrogel composition is administered to an individual suffering from dehydration of the skin, wherein administration of the composition for the skin, thereby treating the dehydration of the skin. 在该实施方案的另一方面,治疗皮肤缺乏弹性的方法包括向患有皮肤缺乏弹性的个体施用本文公开的水凝胶组合物的步骤,其中施用所述组合物增加了皮肤弹性,从而治疗了皮肤缺乏弹性。 In another aspect of this embodiment, a method of treating skin comprising the step of inelastic administering hydrogel composition disclosed herein to the lack of flexibility with skin, wherein administration of the composition increased skin elasticity, thereby treating lack of skin elasticity. 在该实施方案的又一方面,治疗皮肤粗糙的方法包括向患有皮肤粗糙的个体施用本文公开的水凝胶组合物的步骤,其中施用所述组合物减少了皮肤粗糙,从而治疗了皮肤粗糙。 Rough In yet another aspect, the method comprising the step of rough skin suffering hydrogel composition disclosed herein is administered to an individual, wherein administration of the composition reduces skin roughness, thereby treating the skin treatment of rough skin of the embodiment . 在该实施方案的又一方面,治疗皮肤缺乏紧固度的方法包括向患有皮肤缺乏紧固度的个体施用本文公开的水凝胶组合物的步骤,其中施用所述组合物使皮肤更紧,从而治疗了皮肤缺乏紧固度。 In yet another aspect of this embodiment, lack of skin tautness treatment methods include lack of skin tautness administered to an individual suffering from step disclosed herein, a hydrogel composition, wherein said composition is administered to the skin tighter , thereby treating a lack of skin tautness.

[0122] 在该实施方案的又一方面,治疗皮肤拉伸纹或妊娠纹的方法包括向患有皮肤拉伸纹或妊娠纹的个体施用本文公开的水凝胶组合物的步骤,其中施用所述组合物减少或消除了皮肤拉伸纹或妊娠纹,从而治疗了皮肤拉伸纹或妊娠纹。 [0122] In yet another aspect, the hydrogel composition comprising the step of administering to an individual article with skin or stretch marks skin stretch marks disclosed a method of treating stretch marks, or stretch marks of this embodiment, wherein the administration said composition reduces or eliminates skin stretch marks or stretch marks, stretch marks thereby treating the skin or stretch marks. 在该实施方案的另一方面,治疗皮肤苍白的方法包括向患有皮肤苍白的个体施用本文公开的水凝胶组合物的步骤,其中施用所述组合物增强了肤色或光彩,从而治疗了皮肤苍白。 In another aspect, the method comprising the step of having pale skin hydrogel composition disclosed herein is administered to an individual, wherein the composition is administered, or enhance the color brilliance, thereby treating the skin treatment pale skin of this embodiment pale. 在该实施方案的另一方面,治疗皮肤皱纹的方法包括向患有皮肤皱纹的个体施用本文公开的水凝胶组合物的步骤,其中施用所述组合物减少或消除了皮肤皱纹,从而治疗了皮肤皱纹。 In another aspect of this embodiment, a method of treating skin wrinkles comprising the steps herein disclosed hydrogel composition is administered to an individual suffering from skin wrinkles, wherein said composition is administered to reduce or eliminate skin wrinkles, thereby treating skin wrinkles. 在该实施方案的又一方面,治疗皮肤皱纹的方法包括向个体施用本文公开的水凝胶组合物的步骤,其中施用所述组合物使皮肤抗皮肤皱纹,从而治疗了皮肤皱纹。 In another aspect of this embodiment, a method of treating skin wrinkles comprising the steps herein disclosed hydrogel composition is administered to an individual, wherein said composition is administered so that the skin against skin wrinkles, thereby treating skin wrinkles.

[0123] 在一些实施方案中,真皮填充剂具有持续的生物利用率。 [0123] In some embodiments, the dermal filler has sustained bioavailability. 例如,提供了在引入人类皮肤中时,(例如,经皮内或皮下引入人类以修正面部上的软组织孔隙缺损),向人类释放抗坏血酸(或其它维生素)至少约1个月或长达约20个月或更长时间的真皮填充剂。 For example, provided at the introduction into the human skin, (e.g., intradermally or subcutaneously introduction into the human to the pores of soft tissue on the correction surface defect), the release of ascorbic acid (or other vitamins) to a human of at least about 1 month or up to about 20 dermal fillers months or longer.

[0124] 例如,为预测与填充剂持续时间一致的维生素C持续功效,对结合度进行评估。 [0124] For example, consistent with the predicted duration of the filler to sustained efficacy of vitamin C, to assess the degree of binding. 这种评估基于AA2G经醚化与HA结合的制剂。 This assessment based formulations AA2G etherified binding to HA. 所述制剂在生理条件下稳定,但是开始由附着于细胞膜上的葡萄糖苷酶开始释放抗坏血酸(AsA)。 The formulation stable under physiological conditions, but starting from adhering to the cell membrane glucosidase begin to release ascorbic acid (AsA). 由于α-葡萄糖苷酶附着于细胞膜上,所以AsA的释放在填充剂/细胞界面发生。 Since α- glucosidase attached to the cell membrane, so that the release occurs in the filler AsA / cell interface. 进一步地,AsA从HA-AA2G的释放将伴有HA降解以使得AA2G对成纤维细胞可用。 Further, AsA AA2G released from HA-HA degradation will be accompanied by such cells can be used for the fibers to AA2G. 因此AsA的释放取决于HA的AA2G结合度和持续时间。 Thus the release of AsA depends on HA binding AA2G and duration. 结合度为约5mo 1 %的凝胶可在至少长达1个月,例如介于约3〜5个月时期内释放活性维生素C; IOmo 1 % 结合度的凝胶可在至少长达6〜8个月时期内释放活性维生素C;15mol%结合度的凝胶可在至少长达10〜个月时期内释放活性维生素C;30mol%,长达一年半。 Binding of about 5mo 1% gel may be at least as long as one month, for example, between the release of active vitamin C within about 3 ~ 5 month period; IOmo 1% gel of the degree of binding may be up to at least 6 ~ release the active vitamin C during 8 months; gel 15mol% degree of binding may be released within a month period at least as long 10~ active vitamin C; 30mol%, up to a year and a half.

Figure CN108379112AD00241

[0126] *基于凝胶的参数:体积,0 · Icc;浓度,24mg/ml。 [0126] * based on the parameters of the gel: volume, 0 · Icc; concentration, 24mg / ml. (0 · IX 24 X 3 % X 1000) Λ338* 0.1) =2.13 (mM) (0 · IX 24 X 3% X 1000) Λ338 * 0.1) = 2.13 (mM)

[0127] ** 假设: [0127] ** Assumes:

[0128] AsA以恒定速率释放。 [0128] AsA release at a constant rate.

[0129] AsA的有效浓度为0.05mM并且保持有效>2天2.13*2八0.05*30) =2.8(个月) [0129] The effective concentration is 0.05mM and AsA remains valid> 2 days 2.13 0.05 * eight * 2 30) = 2.8 (months)

[0130] 在本发明的一个实施方案中,提供了一种真皮填充剂,其包含与Star-PEG环氧化物交联的透明质酸并且具有以介于约3mol%和约40mol%之间的结合度与透明质酸结合的维生素C衍生物例如,AA2G (抗坏血酸2-葡萄糖苷)、维生食物(3-氨丙基-L-抗坏血酸磷酸酯)和SAP (抗坏血酸磷酸钠)之一。 [0130] In one embodiment of the present invention, there is provided a dermal filler, comprising hyaluronic acid and Star-PEG crosslinked epoxide and having a binding between about 3mol% to about 40mol% vitamin C derivative of hyaluronic acid bound e.g., of AA2G (ascorbic acid 2-glucoside), Vitagen (3-aminopropyl ascorbyl phosphate -L-) and the SAP (sodium ascorbyl phosphate) one.

[0131] 制备这种真皮填充剂的方法包括使季戊四醇缩水甘油醚(Star-PEG环氧化物)与抗坏血酸2-葡萄糖苷(AA2G)按一定比例反应,反应温度和反应时间适合获得含有由4-臂环氧化物加帽的AA2G (AA2G-4臂环氧化物)、未反应的4臂环氧化物和游离AA2G的组合物。 [0131] The method of preparing such dermal fillers include pentaerythritol ether (Star-PEG epoxide) with ascorbic acid 2-glucoside (of AA2G) by a certain percentage, the reaction temperature and reaction time suitable for obtaining containing from 4- armband oxide capped AA2G (AA2G armbands 4-oxide), 4 AA2G armband oxide and free unreacted composition. 4-臂环氧化物加帽的AA2G (AA2G-4臂环氧化物)经环氧基与透明质酸结合。 4- armband oxide capped AA2G (AA2G-4 armband oxide) and epoxy-hyaluronic acid binding. 未反应的4臂环氧化物用作交联剂以交联透明质酸并且用作结合剂以进一步结合AA2G。 4 armband unreacted oxide is used as a crosslinking agent to crosslink the hyaluronic acid binding agent and as further binding AA2G.

[0132] 在本发明的另一实施方案中,提供了一种真皮填充剂,其包含与BDDE交联的透明质酸并且具有以介于约3mol %和约IOmol %之间的结合度与透明质酸结合的维生素C衍生物例如,AA2G (抗坏血酸2-葡萄糖苷)、维生食物(3-氨丙基-L-抗坏血酸磷酸酯)和SAP (抗坏血酸磷酸钠)之一。 [0132] In another embodiment of the present invention, there is provided a dermal filler, comprising hyaluronic acid crosslinked with BDDE and having between about 3mol% to about IOmol% degree of binding to the hyaluronic vitamin C derivative such as an acid binding, of AA2G (ascorbic acid 2-glucoside), Vitagen (3-aminopropyl ascorbyl phosphate -L-) and the SAP (sodium ascorbyl phosphate) one.

[0133] 制备这种真皮填充剂的方法包括使BDDE与抗坏血酸2-葡萄糖苷(AA2G)按一定比例反应,反应温度和反应时间适合获得含有由BDDE加帽的AA2G (AA2G-BDDE)、未反应的BDDE 和游离AA2G的组合物。 [0133] This preparation method comprises contacting dermal fillers BDDE and ascorbic acid 2-glucoside (of AA2G) by a certain percentage, the reaction temperature and reaction time suitable for obtaining a BDDE containing capped AA2G (AA2G-BDDE), unreacted BDDE and free compositions of AA2G. BDDE加帽的AA2G (AA2G-BDDE)经环氧基与透明质酸结合。 BDDE capped AA2G (AA2G-BDDE) epoxy-hyaluronic acid binding. 未反应的BDDE用作交联剂以交联透明质酸并且用作结合剂以进一步结合AA2G。 BDDE as unreacted crosslinking agent used to crosslink the hyaluronic acid binding agent and further binding AA2G.

[0134] 图9是显示α-葡萄糖苷酶浓度对从AA2G-PBS溶液释放AsA的影响的表。 [0134] FIG. 9 is a table α- glucosidase concentration on the release of AsA from the pair AA2G-PBS solution. AA2G向AsA 的转化取决于α_糖苷酶的浓度。 AA2G depends on the concentration of the glycosidase α_ the conversion of AsA. 当α-糖苷酶浓度为6.3个单位/g凝胶时,AA2G在15min内几乎完全转化为AsA。 When α- glycosidase concentration of 6.3 units / g gel, AA2G 15min in almost complete conversion to AsA. 当α-糖苷酶浓度为4.7个单位/g凝胶时,需要30min将AA2G完全转化为AsA。 When α- glycosidase concentration of 4.7 units / g-gel, the required 30min AA2G complete conversion to AsA. 进一步降低α-糖苷酶浓度导致AA2G向AsA的转化缓慢。 To further reduce the concentration of lead α- glucosidase AA2G slow conversion to AsA.

[0135] 图10示出了来自根据本发明所述的结合真皮填充剂的游离AsA的释放曲线示意图(持续释放)(与反应时间相比的AA2G转化mol %)。 [0135] FIG. 10 shows a schematic diagram of the release profile from the free AsA dermal filler binding according to the present invention (sustained release) (as compared with the reaction time AA2G conversion mol%). 在AA2G/HA混合物中AA2G在40min内完全转化为AsA。 In AA2G / HA mixture in 40min AA2G complete conversion to AsA. AA2H/HA结合物显示出AA2G转化为AsA的时间依赖性。 AA2H / HA conjugates exhibits AsA AA2G into time-dependent.

[0136] 图IlA和IlB示出了根据本发明所述的各种真皮填充剂的附加释放数据。 [0136] FIG IlA and IlB release illustrate additional data according to various dermal filler according to the invention. 更具体地,HA-AA2G凝胶中AA2G向AsA的转化取决于α-糖苷酶浓度。 More specifically, HA-AA2G gel AA2G depending α- glycosidase concentration the conversion of AsA. 高的α-糖苷酶浓度导致AA2G向AsA快速转化。 Α- high glycosidase concentration results in rapid conversion of AA2G to AsA. 对于指定α-糖苷酶浓度而言,不同制剂显示出AA2G向AsA转化的不同性质。 For specifying α- glycosidase concentration, different formulations exhibit different properties AA2G conversion to AsA.

[0137] 在本发明的一个方面,提供了真皮填充剂,其对治疗和消除细纹的出现特别有效, 例如皮肤上相对浅表的皱褶,例如但不限于眼睛、泪沟区、前额附近的细纹、眼角纹、眉间纹等。 [0137] In one aspect of the present invention, there is provided a dermal filler, which is particularly effective for the treatment and elimination of occurrence of fine lines, for example, a relatively superficial skin wrinkles, for example, but not limited to the eye, the tear trough area, near the forehead fine lines, eye pattern, frown lines and the like.

[0138] 在已经注射了真皮填充剂的皮肤部位出现蓝色变色(廷德尔效应)是一些真皮填充剂患者经历的重大不良事件。 [0138] In the skin site has been injected with dermal fillers a blue discoloration (Tyndall effect) are some of the dermal filler patients experienced major adverse events. 廷德尔效应在治疗浅表细小皱纹的患者中更常见。 Tyndall effect is more common in patients in the treatment of superficial fine wrinkles. 已经研发了本发明的实施方案,其提供了可在浅表注射以治疗细纹和皱纹,甚至在相对较薄的皮肤区域中注射,没有任何由廷德尔效应产生的蓝色变色的长效、半透明填充剂。 Have been developed embodiment of the present invention, which provides a superficial injection to treat wrinkles and fine lines, even in the injection area of ​​the skin is relatively thin, there is no blue Tyndall effect resulting from long-term discoloration, translucent filler. 细纹或浅表皱纹一般被理解为通常在皮肤最薄,即皮肤的真皮厚度小于Imm的面部区域(前额、外眦、唇红缘/口周线)中发现的皮肤上的皱纹或皱褶。 Superficial wrinkles or fine lines as is generally understood in the skin is generally thinnest, i.e. less than the thickness of the skin dermal wrinkles or folds on the discovery (forehead, canthus, vermilion border / perioral lines) of the face region of the skin Imm . 在前额,平均真皮厚度对于正常皮肤而言为约0.95mm而对于起皱皮肤而言为约0.8lmmm。 The forehead, the average for normal skin dermal thickness of about 0.95mm and wrinkled skin for about 0.8lmmm. 外目此周围的真皮甚至更薄(例如对于正常皮肤而言约0.61mm而对于起皱皮肤而言约0.41mm)。 This mesh around the outer leather even thinner (e.g. about 0.61mm for normal skin and skin wrinkles for about 0.41mm). 30或32G针(通常用于细纹凝胶应用的针)的平均外径为约〇. 30和约0.24mm。 The average outer diameter (typically a needle for application of the gel fine lines) or the 32G needle was about 30 billion. 30 and about 0.24mm.

[0139] 本发明提供了例如在本文其它地方描述的不会导致廷德尔效应的真皮填充剂组合物。 [0139] The present invention provides, for example, will not described elsewhere herein causes dermal filler composition of the Tyndall effect. 例如,本发明的组合物包含与交联剂交联的透明质酸组分、除交联组分外的添加剂; 当施用到患者的真皮区内时,相对于除没有所述添加剂外,大体上相同的组合物,所述组合物表现出廷德尔效应减小。 For example, compositions of the present invention comprises a crosslinking agent with hyaluronic acid component, in addition to the crosslinking additive exceptionally group; when administered to a patient dermal region, without addition of the additive with respect to an outer, generally the same composition, the composition exhibits reduced Tyndall effect. 所述组合物可大体上光学透明。 The composition may be substantially optically transparent.

[0140] 在一个实施方案中,添加剂是维生素C衍生物,例如可与本文其它地方描述的透明质酸化学结合的AA2G。 [0140] In one embodiment, the additive is a vitamin C derivative, for example, hyaluronic acid chemically described elsewhere herein, binding of AA2G.

[0141] 在一些实施方案中,交联组分是BDDE并且结合度介于约3mol %和约IOmol %之间, 或高达15mol%或更高。 [0141] In some embodiments, the crosslinking component is BDDE and binding of between about 3mol% and about IOmol%, or up to 15mol% or more. 在一些实施方案中,所述组合物进一步包含适于在注射后为患者提供舒适的量的麻醉剂,例如利多卡因。 In some embodiments, the composition further comprises an anesthetic in an amount suitable for providing comfort for the patient after injection, such as lidocaine.

[0142] 还提供了治疗患者皮肤上的细纹的方法。 [0142] Also provided are methods of treating fine lines on the skin of a patient. 所述方法通常包括向患者皮肤内引入例如本文所述的组合物的步骤。 The method generally includes the step of, for example, a composition described herein is introduced into the patient's skin. 例如所述组合物包含透明质酸组分、交联透明质酸的交联组分和除交联组分外的添加剂的混合物,所述组合物大体上光学透明;并且其中相对于除没有所述添加剂外,大体上相同的组合物,真皮填充剂组合物表现出廷德尔效应减小。 For example, the composition comprising hyaluronic acid component, a crosslinked hyaluronic acid and a crosslinking component in addition to the components of the crosslinking additive mixture, the composition is substantially optically transparent; and wherein no other relative to the said external additive, substantially the same composition, dermal filler composition exhibits reduced Tyndall effect.

[0143] 在本发明的一些实施方案中,所述组合物包含使用EDC化学与二胺或多胺交联剂交联的透明质酸组分。 [0143] In some embodiments of the present invention, the composition comprising hyaluronic acid and a diamine component using EDC or polyamine chemical crosslink. 例如,所述交联剂可为HMDA。 For example, the crosslinking agent may be HMDA.

[0144] 在交联剂为HMDA的本发明的某些实施方案中,所述组合物的G'高达约70Pa,G”/G' 介于约0.65和约0.75之间,挤压力约24N或更小,并且最终HA浓度介于约24mg/g和约25mg/g 之间。 [0144] Certain embodiments of the present invention is in the crosslinking agent HMDA, said composition G 'up to about 70Pa, G "/ G' is between about 0.65 and about 0.75, the pressing force of about 24N or It is smaller, and the final HA concentration is between about 24mg / g and about 25mg / g.

[0145] 在添加剂为HA-AA2G结合物或HA-维生食物结合物的本发明的某些实施方案中,结合度介于约3111〇1%和约1〇1]1〇1%之间,或高达约151]1〇1%,或高达约4〇1]1〇1%。 Certain embodiments of the invention [0145] The conjugate or conjugate Vitagen HA- HA-AA2G as additives in the binding of between about% and about 1〇1 3111〇1] 1〇1% or up to about 151] 1〇1%, or up to about 4〇1] 1〇1%. 这些组合物的G'从至少约30Pa,更优选至少约40Pa到约IOOPa,G” /G'介于约0.30和约0.50之间,挤压力约27N或更小,并且最终HA浓度介于约24mg/g和约25mg/g之间。 These compositions G 'of from at least about 30 Pa, more preferably at least about 40Pa to about IOOPa, G "/ G' is between about 0.30 and about 0.50, the pressing force of about 27N or less, and the final HA concentration of between about 24mg / g between about 25mg / g.

[0146] 为了本发明的目的,如本文所使用的“结合度”定义为结合体的摩尔百分比,例如AA2G与透明质酸重复单元(例如,HA二聚体)。 [0146] For purposes of the present invention, such as "binding" is defined herein as the molar percentage of the combination, e.g. AA2G hyaluronic acid repeating unit (e.g., the HA dimer). 因此,IOmol %结合度意指每100个HA重复单元含10个结合的AA2G。 Thus, IOmol% means that binding of HA repeating units per 100 containing 10 binding AA2G. 可使用下面实施例2中说明的方法,或本领域技术人员已知的其它方法计算结合度。 2 may be used in the following examples illustrate embodiments of a method, or other methods known to those skilled in the degree of binding is calculated. 实施例 Example

[0147] 实施例1:使用BDDE作为交联剂AA2G与交联的HA凝胶结合 [0147] Example 1: Use as a crosslinking agent BDDE AA2G crosslinked HA gel with binding

[0148] 使400.6mg的低分子量透明质酸(LMW HA)在注射器中1802mg的Iwt^NaOH中水合〜30min。 [0148] 400.6mg of low molecular weight hyaluronic acid (LMW HA) Iwt 1802mg ^ NaOH in a syringe hydrated ~30min. 将800 · 7mg的AA2G放入小瓶中,接着放入713 · 7mg的BDDE和1416 · 8mg的10%Na0H。 The 800 · 7mg AA2G is put in a vial, and then placed in 10% Na0H 713 · 7mg of BDDE and 1416 · 8mg of. 在添加到水合HA中之前,使以上溶液(pH>12)在50°C水浴中反应〜20min。 Prior to addition to the hydrated HA, the above solution (pH> 12) 50 ° C and the reaction ~20min water bath. 添加之后,通过在2个注射器之间来回传递使混合物混合〜20次。 After the addition, by passing back and forth between the two syringes mixed and the mixture was ~ 20 times. 将混合的糊剂放入小瓶和50°C水浴中〜 2.5h。 The mixed paste was put in a vial and 50 ° C water bath for ~ 2.5h. 将223.5mg的12M HCl添加到9.05g PBS(pH7.4)中。 223.5mg of 12M HCl added to 9.05g PBS (pH7.4) in. 〜2.5h后,形成HA-AA2G凝胶。 After ~2.5h, HA-AA2G gel is formed. 将凝胶切成片,并且向其中添加HCl-PBS溶液。 The gel was cut into pieces, and a solution of HCl-PBS was added thereto. 使凝胶中和并且在定轨振荡器上膨胀整夜。 The gel was neutralized and expanded on an orbital shaker overnight. 通过〜60μπι筛子筛分凝胶并且通过在2个注射器之间来回传递混合〜20次。 ~60μπι sieved through sieve gel and transferred back and forth between two syringes through a mixing ~ 20 times. 将凝胶放入15,000 MffCO RC透析袋中并且在PBS (ρΗ7.4)缓冲液中透析。 The gel was put 15,000 MffCO RC dialysis bag and dialyzed in PBS (ρΗ7.4) buffer. 透析进行〜185h,频繁更换PBS缓冲液。 Dialysis ~185h, frequent change of PBS buffer. 透析后,将凝胶放入注射器中并且储存在4°C冰箱中。 After dialysis, the gel was placed in a syringe and stored at 4 ° C refrigerator.

[0149] 实施例2:AA2G结合的测定 AA2G binding assay: two cases of [0149] Embodiment

[0150] 透析之前和透析之后正确记录如实施例1所述的凝胶的重量。 [0150] The correct recording of the weight of the gel in Example 1 before and after dialysis and dialysis. 假设透析之后凝胶为〜lg/mL。 After dialysis gel is assumed ~lg / mL. 在IL PBS中每>8h,没有出现显而易见的AA2G时停止透析。 Each> 8h, stop dialysis apparent AA2G does not appear in the IL PBS. 使用UVVis分光光度计(Nanodrop 2000C,ThermoScientif ic)在260nm下测量AA2G。 UVVis using a spectrophotometer (Nanodrop 2000C, ThermoScientif ic) AA2G measured at 260nm. 使用2%HA中AA2G的不同浓度(A@260nm= 1 · 4838 [AA2G (mM)])计算AA2G的校准曲线。 AA2G using 2% HA in different concentrations (A @ 260nm = 1 · 4838 [AA2G (mM)]) of the calculation of the calibration curve AA2G.

[0151] 透析之后HA的重量:HA的起始重量X (透析之前的实际重量/理论重量) [0151] HA weight after dialysis: HA starting weight X (weight before dialysis actual / theoretical weight)

[0152] 透析之后AA2G的mmol:透析之后在方程式(A@260nm=l .4838 [AA2G (mM)])中将吸收设于260nm下。 [0152] mmol of AA2G after dialysis: after dialysis in Equation (A 260nm @ = l .4838 [AA2G (mM)]) provided in the absorption at 260nm.

[0153] AA2G的结合(AA2G的mmol/mmol的HA) X100% Binding [0153] of AA2G (mmol / mmol of AA2G of HA) X100%

[0154] 如实施例1中所述凝胶中的AA2G结合度为14.7mol %。 [0154] The binding of AA2G in Example 1. The gel was 14.7mol%.

[0155] 实施例3:凝胶流变性的测定 3 [0155] Example: Measurement of Gel Rheology

[0156] 使用振荡平行板流变仪(Anton Paar,Physica MCR 301)测量实施例1中获得的凝胶的性质。 Properties of the gel obtained in Example 1 [0156] Using the oscillating parallel plate rheometer (Anton Paar, Physica MCR 301) measured embodiments. 所用板的直径为25mm。 The plate diameter is 25mm. 板间间隙设为1mm。 Gap between the plates to 1mm. 对于每次测量而言,在固定频率下应变扫描之前,首先进行在恒定应变下的频率扫描。 For each measurement, the strain before scanning at a fixed frequency, first frequency sweep under constant strain. 在1%应变下由应变扫描曲线获得G'(储能模量)。 Obtained at 1% strain G '(storage modulus) from the strain sweep curves. 对凝胶而言值为1450Pa。 For gel is 1450Pa.

[0157] 实施例4:使用BDDE作为交联剂,以可调结合度和凝胶流变性AA2G与交联的HA凝胶结合 [0157] Example 4: BDDE used as a crosslinking agent, in combination with an adjustable degree and gel rheological AA2G crosslinked HA gel with binding

[0158] 程序与实施例1中描述的相似。 [0158] procedures and embodiments similar to that described in Example 1. 通过调整交联剂与HA和AA2G摩尔比更改结合度。 The molar ratio of crosslinking agent to HA and AA2G changed by adjusting the degree of binding. 如实施例3中所述测量凝胶性质。 As in Example 3. The measured properties of the gel. 详情如下: Details are as follows:

[0159] 使400.8mg的LMff HA在注射器中1752. Img的I %NaOH中水合〜30min。 [0159] In so LMff HA 400.8mg of syringe 1752. Img I% NaOH hydrated ~30min. 将800.3mg的AA2G放入小瓶中,接着放入354 .Img的BDDE和1402 .Omg的10%NaOH。 The 800.3mg of AA2G put in a vial and then placed in a 354 .Img BDDE and 1402 .Omg of 10% NaOH. 在添加到水合HA中之前,使以上溶液(pH>12)在50°C水浴中反应〜20min。 Prior to addition to the hydrated HA, the above solution (pH> 12) 50 ° C and the reaction ~20min water bath. 添加之后,通过在2个注射器之间来回传递使混合物混合〜20次。 After the addition, by passing back and forth between the two syringes mixed and the mixture was ~ 20 times. 将混合的糊剂放入小瓶和50°C水浴中〜2.5h。 The mixed paste was put in a vial and 50 ° C water bath ~2.5h. 将140.9mg的12M HCl添加到9.0053g PBS (pH7.4)中。 140.9mg of 12M HCl added to 9.0053g PBS (pH7.4) in. 〜2.5h后,形成HA-AA2G凝胶。 After ~2.5h, HA-AA2G gel is formed. 将凝胶切成片,并且向其中添加HCl-PBS溶液。 The gel was cut into pieces, and a solution of HCl-PBS was added thereto. 使凝胶中和并且在定轨振荡器上膨胀整夜。 The gel was neutralized and expanded on an orbital shaker overnight. 通过〜60μπι筛子筛分凝胶并且通过在2个注射器之间来回传递混合〜20次。 ~60μπι sieved through sieve gel and transferred back and forth between two syringes through a mixing ~ 20 times. 将凝胶放入15,000MWC0 RC透析袋中并且在PBS (pH7.4)缓冲液中透析。 The gel was placed in a dialysis bag and dialyzed 15,000MWC0 RC in PBS (pH7.4) buffer. 透析进行〜164.5h,频繁更换PBS缓冲液。 Dialysis ~164.5h, frequent change of PBS buffer. 透析后,将凝胶放入注射器中并且储存在4°C冰箱中。 After dialysis, the gel was placed in a syringe and stored at 4 ° C refrigerator. 结合度为13%。 13% degree of binding. 凝胶储能模量(G')为803Pa。 Gel storage modulus (G ') of 803Pa.

[0160] 实施例5:使用BDDE作为交联剂,AA2G与交联的HA凝胶结合,结合度为5.3%,G'为〜300Pa〇 [0160] Example 5: BDDE used as a crosslinking agent, of AA2G crosslinked HA gel with binding, binding degree is 5.3%, G 'is ~300Pa〇

[0161] 使400.3mg的LMff HA在注射器中3002. Omg的I %NaOH中水合〜30min。 [0161] 400.3mg of making LMff HA in the syringe 3002. I% NaOH Omg hydrated ~30min. 将800.5mg的AA2G放入小瓶中,接着放入264 · 3mg的BDDE和1100 · Omg的10 %NaOH。 The 800.5mg of AA2G put in a vial and then placed in a 264 · 3mg BDDE and 1100 · Omg of 10% NaOH. 在添加到水合HA中之前,使以上溶液(pH>12)在50°C水浴中反应〜20min。 Prior to addition to the hydrated HA, the above solution (pH> 12) 50 ° C and the reaction ~20min water bath. 添加之后,通过在2个注射器之间来回传递使混合物混合〜20次。 After the addition, by passing back and forth between the two syringes mixed and the mixture was ~ 20 times. 将混合的糊剂放入小瓶和50°C水浴中〜2.5h。 The mixed paste was put in a vial and 50 ° C water bath ~2.5h. 将104.2mg的12M HCl添加到8.5128g PBS (pH7.4)中。 104.2mg of 12M HCl added to 8.5128g PBS (pH7.4) in. 〜2.5h后,形成HA-AA2G凝胶,并且向其中添加HCl-PBS 溶液。 After ~2.5h, HA-AA2G gel is formed, and a solution of HCl-PBS was added thereto. 使凝胶中和并且在定轨振荡器上膨胀整个周末(〜55h)。 The gel was neutralized and expanded over the weekend (~55h) on an orbital shaker. 通过〜60μπι筛子筛分凝胶并且通过在2个注射器之间来回传递混合〜20次。 ~60μπι sieved through sieve gel and transferred back and forth between two syringes through a mixing ~ 20 times. 将凝胶放入15,000 MWCO RC透析袋中并且在PBS (ρΗ7.4)缓冲液中透析。 The gel was placed in 15,000 MWCO RC dialysis bag and dialyzed in PBS (ρΗ7.4) buffer. 透析进行〜114h,频繁更换PBS缓冲液。 Dialysis ~114h, frequent change of PBS buffer. 透析后,将凝胶放入注射器中并且储存在4°C冰箱中。 After dialysis, the gel was placed in a syringe and stored at 4 ° C refrigerator. 按实施例2和3中描述的程序测量结合度和凝胶流变性。 Procedure described in Example 2 and 3 in conjunction with measurement of rheological and gel. 结合度为5.3%。 Binding of 5.3%. 凝胶储能模量为〜300卩&。 Gel storage modulus Jie ~ 300 & amp ;.

[0162] 实施例6:使用star-PEG环氧化物作为交联剂,AA2G与交联的HA凝胶结合,结合度为29.4%,G' 为〜235Pa。 [0162] Example 6: Use star-PEG epoxide as a crosslinking agent, of AA2G crosslinked HA gel with binding, binding degree is 29.4%, G 'is ~235Pa.

[0163] 使200.4mg的LMff HA在注射器中2000mg的l%NaOH中水合〜30min。 [0163] 200.4mg of LMff HA 2000mg so in a syringe l% NaOH hydrated ~30min. 将400mg的AA2G 放入小瓶中,接着放入312.7mg的star-PEG环氧化物和1026.5mg的10%Na0H。 The 400mg of AA2G put in a vial and then placed in star-PEG epoxide and 10% Na0H 1026.5mg of 312.7mg of. 在添加到水合HA中之前,使以上溶液在50 °C水浴中反应〜20min。 Prior to addition of the hydrated HA to the reaction solution above 50 ° C and ~20min water bath. 添加之后,通过在2个注射器之间来回传递使混合物混合〜2 0次。 After the addition, by passing back and forth between two syringes and the mixture was mixed ~ 2 0 times. 将混合的糊剂放入小瓶和5 0 °C水浴中〜2.5 h。 The mixed paste was put in a vial and 5 0 ° C water bath ~2.5 h. 将18 7.4 mg的12 M HCl添加到3 · 034g I3BS (pH7 · 4)中。 Adding 18 7.4 mg of 12 M HCl to 3 · 034g I3BS (pH7 · 4) in. 〜2 · 5h后,形成HA-AA2G凝胶,并向其中添加HCl-I3BS溶液。 After ~2 · 5h, HA-AA2G gel is formed, and wherein the solution was added HCl-I3BS. 使凝胶中和并且在定轨振荡器上膨胀整个周末(〜68h)。 The gel was neutralized and expanded over the weekend (~68h) on an orbital shaker. 通过〜60μπι筛子筛分凝胶并且通过在2个注射器之间来回传递混合〜20次。 ~60μπι sieved through sieve gel and transferred back and forth between two syringes through a mixing ~ 20 times. 将凝胶放入15,000MW⑶RC透析袋中并且在PBS (ρΗ7.4)缓冲液中透析。 The gel was put 15,000MW⑶RC dialysis bag and dialyzed in a buffer PBS (ρΗ7.4). 透析进行〜95h,频繁更换PBS缓冲液。 Dialysis ~95h, frequent change of PBS buffer. 透析后,将凝胶放入注射器中并且储存在4 °C冰箱中。 After dialysis, the gel was placed in a syringe and stored at 4 ° C refrigerator. 按实施例2和3中描述的程序测量结合度和凝胶流变性。 Procedure described in Example 2 and 3 in conjunction with measurement of rheological and gel. 结合度为29.4%。 Binding of 29.4%. 凝胶储能模量为〜235Pa。 Gel storage modulus ~235Pa.

[0164] 实施例7:使用star-PEG环氧化物作为交联剂,AA2G与交联的HA凝胶结合,结合度为27.8%,G' 为〜363Pa。 [0164] Example 7: Use star-PEG epoxide as a crosslinking agent, of AA2G crosslinked HA gel with binding, binding degree is 27.8%, G 'is ~363Pa.

[0165] 使200 · 3mg的LMW HA在注射器中2000mg的I %NaOH中水合〜30min。 [0165] LMW HA 200 · 3mg so in a syringe I% NaOH ~30min 2000mg of hydration. 将400 · 2mg的AA2G放入小瓶中,接着放入313.4mg的star-PEG环氧化物和1022.6mg的10%NaOH。 The 400 · 2mg AA2G is put in a vial and then placed in a star-PEG 313.4mg epoxide and 1022.6mg of 10% NaOH. 将以上溶液添加到水合HA中。 The above solution was added to the hydrated HA. 添加之后,通过在2个注射器之间来回传递使混合物混合〜20次。 After the addition, by passing back and forth between the two syringes mixed and the mixture was ~ 20 times. 将混合的糊剂放入小瓶和50°C水浴中〜2 · 5h。 The mixed paste was put in a vial in a water bath at 50 ° C and ~2 · 5h. 将196 · 5mg的12M HCl添加到3 · 016g PBS (pH7 · 4) 中。 Adding 196 · 5mg of 12M HCl to 3 · 016g PBS (pH7 · 4) in. 〜2.5h后,形成HA-AA2G凝胶,并向其中添加HCl-PBS溶液。 After ~2.5h, HA-AA2G gel is formed, and wherein the solution was added HCl-PBS. 使凝胶中和并且在定轨振荡器上膨胀整夜(〜24h)。 The gel was neutralized and expanded on an orbital shaker overnight (~24h). 通过〜60μπι筛子筛分凝胶并且通过在2个注射器之间来回传递混合〜20次。 ~60μπι sieved through sieve gel and transferred back and forth between two syringes through a mixing ~ 20 times. 将凝胶放入15,000 MWCO RC透析袋中并且在PBS (ρΗ7.4)缓冲液中透析。 The gel was placed in 15,000 MWCO RC dialysis bag and dialyzed in PBS (ρΗ7.4) buffer. 透析进行〜98.5h,频繁更换PBS缓冲液。 Dialysis ~98.5h, frequent change of PBS buffer. 透析后,将凝胶放入注射器中并且储存在4°C冰箱中。 After dialysis, the gel was placed in a syringe and stored at 4 ° C refrigerator. 按实施例2和3中描述的程序测量结合度和凝胶流变性。 Procedure described in Example 2 and 3 in conjunction with measurement of rheological and gel. 结合度为27.8 %。 Binding of 27.8%. 凝胶储能模量为〜 363Pa〇 Gel storage modulus ~ 363Pa〇

[0166] 实施例8:使用BDDE作为交联剂,AA2G与交联的HMW HA凝胶结合,结合度为约10mol%,G' 为约240Pa。 [0166] Example 8: Use as a crosslinking agent BDDE, of AA2G crosslinked gel with HMW HA binding, binding of about 10mol%, G 'of about 240Pa.

[0167] 使400.3mg的HMW HA在注射器中2501.3mg的4wt%NaOH中水合〜30min。 [0167] that the HMW HA 400.3mg hydrated in a syringe ~30min 4wt% NaOH 2501.3mg of. 将1200mg 的AA2G放入小瓶中,接着放入304 · 7mg的BDDE和1178 · 6mg的16wt%NaOH。 The 1200mg of AA2G put in a vial and then placed in a 16wt% 304 · 7mg BDDE and 1178 · 6mg of NaOH. 在添加到水合HA之前,使以上溶液(pH>12)在50°C水浴中反应〜20min。 Prior to addition to the hydrated HA, the above solution (pH> 12) 50 ° C and the reaction ~20min water bath. 添加之后,通过在2个注射器之间来回传递使混合物混合〜20次。 After the addition, by passing back and forth between the two syringes mixed and the mixture was ~ 20 times. 将混合的糊剂放入20cc小瓶和50°C水浴中〜2.5h。 The mixed paste into 20cc vials and 50 ° C water bath ~2.5h. 〜2.5h后,形成HA-AA2G凝胶。 After ~2.5h, HA-AA2G gel is formed. 将226.6mg的12M HCl添加到8492.2mg 10XPBS(pH7.4)中获得HCl-PBS溶液并且添加HCl-PBS溶液以使凝胶中和和膨胀。 Adding 226.6mg 8492.2mg 10XPBS of 12M HCl to obtain a solution of HCl-PBS (pH7.4) and added HCl-PBS solution was added to neutralize and swell the gel. 使凝胶中和并且在定轨振荡器上膨胀48h以上。 The gel was neutralized and expanded for 48h on an orbital shaker. 通过〜60μπι筛子筛分凝胶并且通过在2个注射器之间来回传递混合〜20次。 ~60μπι sieved through sieve gel and transferred back and forth between two syringes through a mixing ~ 20 times. 将凝胶放入20,000 MffCO CE透析袋中并且在PBS (ρΗ7.4)缓冲液中透析。 The gel was put 20,000 MffCO CE dialysis bag and dialyzed in PBS (ρΗ7.4) buffer. 透析进行〜114h,频繁更换PBS 缓冲液。 Dialysis ~114h, frequent change of PBS buffer. 透析后,将凝胶放入注射器中并且储存在4°C冰箱中。 After dialysis, the gel was placed in a syringe and stored at 4 ° C refrigerator. 按实施例2和3中描述的程序测量结合度和凝胶流变性。 Procedure described in Example 2 and 3 in conjunction with measurement of rheological and gel. 结合度为IOmol %。 Degree of binding IOmol%. 凝胶储能模量为约240Pa。 Gel storage modulus of about 240Pa.

[0168] 实施例9:使用BDDE作为交联剂,维生食物与交联的LMW HA凝胶结合,结合度为15mol%,G' 为约365Pa。 [0168] Example 9: BDDE used as a crosslinking agent, in combination with Vitagen LMW HA crosslinked gel, binding degree of 15mol%, G 'of about 365Pa.

[0169] 使398.2mg的LMff HA在注射器中1753.24mg的Iwt%NaOH中水合〜40min。 [0169] 398.2mg of LMff HA 1753.24mg so in a syringe Iwt% NaOH hydrated ~40min. 向膨胀的HA中添加BDDE (311.7mg)并且继续让HA再膨胀80min。 Add BDDE (311.7mg) to continue the expansion of the HA and HA allow re-expansion 80min. 使膨胀的HA/BDDE混合物在50 °C下预先反应20min。 Swollen HA / BDDE at 20min the reaction mixture was pre-50 ° C.

[0170] 将801.9mg的维生食物单独溶于1459.7mg的IOwt %NaOH中并且和与BDDE预先反应的HA混合。 [0170] 801.9mg of the dissolved HA alone Vitagen mixing IOwt% NaOH 1459.7mg of BDDE and and a pre-reacted. 使混合物继续在50 °C下再反应2.5h。 The mixture was further continued to react at 50 ° C 2.5h. 〜2.5h后,形成HA-维生食物。 After ~2.5h, form HA- Vitagen. 将195mg的12M HCl添加到9004. Omg的10 X PBS (pH7.4)中获得HCl-PBS溶液并且添加HCl-I3BS溶液以使凝胶中和和膨胀。 Adding to 195mg of 12M HCl 9004. Omg of 10 X PBS (pH7.4) was obtained in HCl-PBS and added HCl-I3BS solution was added to neutralize and swell the gel. 使凝胶中和并且在定轨振荡器上膨胀48h以上。 The gel was neutralized and expanded for 48h on an orbital shaker. 通过〜60μπι筛子筛分凝胶并且通过在2个注射器之间来回传递混合〜20次。 ~60μπι sieved through sieve gel and transferred back and forth between two syringes through a mixing ~ 20 times. 将凝胶放入20,000MW⑶CE透析袋中并且在PBS (pH7.4)缓冲液中透析。 The gel was placed in a dialysis bag and dialyzed 20,000MW⑶CE in PBS (pH7.4) buffer. 透析进行〜120h,频繁更换PBS缓冲液。 Dialysis ~120h, frequent change of PBS buffer. 透析后,将凝胶放入注射器中并且储存在4°C冰箱中。 After dialysis, the gel was placed in a syringe and stored at 4 ° C refrigerator. 按实施例3中描述的程序测量凝胶流变性。 Gel Rheology measurements according to the procedure described in Example 3. 使用与实施例2中描述的AA2G测定相似的方法测定结合度为约15mol %。 Determination of binding of about 15mol% using a similar method AA2G assay described in Example 2. 凝胶储能模量为约365Pa。 Gel storage modulus of about 365Pa.

[0171] 实施例10:维生食物经酰胺化学与线性HA结合 [0171] Example 10: Vitagen HA binding by linear chemistry

[0172] 使200.3mg的HMW HA在60cc注射器中IOml的水中水合。 [0172] that the HMW HA 200.3mg IOml hydration water in 60cc syringe. 将500mg的维生食物溶于0.5ml水中并且将溶液中和到pH 4.8。 500mg of the Vitagen dissolved in 0.5ml of water and the solution neutralized to pH 4.8. 将197.7mg的EDC和149mg的NHS单独溶于6ml水中。 The 149mg of EDC and NHS 197.7mg separately dissolved in 6ml water. 将以上溶液(溶液和H)C/NHS溶液)添加到另一装有23.5ml水的60cc注射器中。 The above solution (solution H) C / NHS solution) was added to a 60cc syringe with another 23.5ml water. 通过在2个注射器之间来回传递混合20次。 Mixing by passing 20 times back and forth between two syringes. 将混合物储存在一个注射器中并且浸入37°C浴中4h。 The mixture was stored in a syringe and immersed in a bath at 37 ° C 4h. 最后用PBS (pH7.4)缓冲液透析溶液,直至没有观察到显而易见的维生食物。 Finally, PBS (pH7.4) buffer solution, dialyzed, was not observed until Vitagen apparent. 通过如实施例3描述的类似方法测定结合度。 Determination binding by analogous methods as described in Example 3. 结合度为约IOmol %。 Binding of about IOmol%.

[0173] 实施例11 :AA2P与交联的HA凝胶的结合 Conjunction with HA gel of crosslinked AA2P: [0173] Example 11

[0174] 使200.4mg的LMff HA在注射器中IOOOmg的MES 5.2缓冲液中水合〜3〇111丨11。 [0174] 200.4mg of LMff HA IOOOmg so in a syringe MES 5.2 ~3〇111 Shu hydrated buffer 11. 将29211^ 的AA2P放入小瓶,接着添加300mg的star-PEG胺。 29211 ^ The AA2P placed in a vial, followed by addition of 300mg of star-PEG amine. 使以上溶液在室温下反应整夜。 The reaction above solution at room temperature overnight. 用PBS缓冲液使凝胶水合并且用PBS缓冲液透析以去除未反应的AA2P。 With PBS buffer and the gel hydration dialyzed with PBS buffer to remove unreacted AA2P. 如实施例2和3所述,表征最后的凝胶以测定结合度和凝胶流变性。 As described in Example 2 and 3, a final characterization to determine binding of the gel and gel rheology. 结合度为约20mol%。 Binding of about 20mol%. 储能模量(G')为约500Pa。 Storage modulus (G ') of about 500Pa.

[0175] 实施例12 [0175] Example 12

[0176] 用于减少细纹出现的具有AA2G的HA/BDDE真皮填充剂产品的配制 [0176] formulation for reducing the appearance of fine lines having the HA of AA2G / BDDE dermal filler product

[0177] 对以上实施例中描述的任何凝胶,在透析后,向凝胶中添加适量游离HA凝胶以提高改变凝胶粘聚性和/或可注射性。 [0177] Any of the gel described in the above embodiment, after dialysis, adding an appropriate amount of free HA gel to the gel to enhance the cohesiveness of the gel change, and / or injectable. 例如,使游离HA纤维在磷酸盐缓冲液中膨胀,以便获得均匀粘弹性凝胶(“游离”HA凝胶)。 For example, the free HA fibers swell in phosphate buffer in order to obtain a homogeneous viscoelastic gel ( "free" HA gel). 在透析步骤之前,将这种未交联的凝胶添加到实施例1中获得的HA/BDDE交联凝胶中(例如,以获得具有约1%至约5%w/w游离HA的组合物)。 Before the dialysis step, which would add to the non-crosslinked HA gel obtained in Example 1 of the embodiment / BDDE crosslinked gel (e.g., to obtain a composition having from about 1% to about 5% w / w of free HA matter). 然后将所得凝胶填充到易于填充的无菌注射器中并且在足够温度和压力下高压蒸汽灭菌至少约lmin。 The resulting gel is then filled into a sterile syringe filled easily and at a sufficient temperature and high pressure steam sterilization at least about lmin. 高压蒸汽灭菌后,包装HA/AA2G最终产品并分发给医师以用作真皮填充剂浅表注射, 改善眶周或其它面部区域中细纹的出现。 After autoclaving, packaging HA / AA2G final product and distributed to physicians as superficial dermal filler injection, appears to improve the periorbital lines or other areas of the face.

[0178] 实施例13 [0178] Example 13

[0179] 包括利多卡因的HA-AA2G真皮填充剂的配制 [0179] HA-AA2G comprising lidocaine dermal filler formulation

[0180] 按照实施例12的程序,但是在透析步骤之后和添加游离HA凝胶之前,向混合物中添加利多卡因盐酸盐(利多卡因HC1)。 [0180] Following the procedure of Example 12, but after the dialysis step and before adding the free HA gel was added lidocaine hydrochloride (HC1 is lidocaine) to the mixture. 首先将粉末形式的(利多卡因HC1)溶解于WFI中并且通过0.2μπι过滤器过滤。 First, in powder form (lidocaine HC1 is) is dissolved in WFI and filtered through a filter 0.2μπι. 向粘性HA/AA2G凝胶中添加NaOH稀溶液以便达到弱碱性pH (例如,介于约7.5和约8之间的pH)。 AA2G gel was added a solution of NaOH dilute viscous HA / weakly basic to reach pH (e.g., pH between about 7.5 and about 8). 然后将利多卡因HCl溶液添加到弱碱性凝胶中以达到最终所需浓度,例如约0.3% (w/w)的浓度。 Lidocaine HCl solution was then added to a concentration of weakly basic gel to achieve a final desired concentration, for example about 0.3% (w / w) of. 然后HA/AA2G/利多卡因混合物的所得pH为约7而HA浓度为约24mg/g。 Then HA / AA2G / lidocaine resulting pH of the mixture to about 7 and the HA concentration of about 24mg / g. 在装备有适当搅拌机的标准反应器中进行机械混合以便获得适当均匀性。 In a standard reactor equipped with a suitable mechanical stirrer to obtain an appropriate mixing uniformity.

[0181] 实施例14 [0181] Example 14

[0182] 含羧基官能团的添加剂与HA水凝胶的结合 [0182] Additives containing carboxyl functional group binding to the HA hydrogel

[0183] 添加剂,例如视黄酸(AKA,维甲酸)、阿达帕林(adapalence)和α-硫辛酸含有羧基官能团(-CO⑽。使用EDC化学经酯化作用,使这些添加剂与HA水凝胶结合。以下描述了根据本发明一个实施方案所述的结合的实例: [0183] Additives, such as retinoic acid (the AKA, tretinoin), adapalene (adapalence) and α- lipoic acid containing carboxyl functional group (-CO⑽. By esterification using EDC chemistry, these additives HA hydrogels combination examples described below, according to one embodiment of the present invention bound:

[0184] 使200mg的HMW HA在60cc注射器中IOml的pH 4.8MES缓冲液中水合。 [0184] so that the HMW HA 200mg 4.8MES buffer pH IOml hydrated in the 60cc syringe. 在另一个注射器中,将200mg的视黄酸溶于5ml的水-丙酮混合物(水/丙酮体积比1:3)。 In another syringe, 200mg of the retinoic acid was dissolved in 5ml of water - acetone mixture (water / acetone volume ratio of 1: 3). 通过注射器连接器混合以上两个注射器约20次。 Mixing about 20 times more than the two syringes via a syringe connector. 然后将197.7mg的EDC和149mg的NHS分别溶于单独注射器中的6ml水中。 Then 149mg of EDC and NHS 197.7mg were dissolved in 6ml water in separate syringes. 装有EDC和NHS的注射器与装有HA和视黄酸的注射器连接,以通过在2个注射器之间来回传递使反应物混合至少20次。 With EDC and NHS syringe connected to the syringe containing the HA and retinoic acid, through the back and forth between the two syringes pass the reaction mixture was at least 20 times. 将混合物储存在一个注射器中并且浸入37°C浴中4h。 The mixture was stored in a syringe and immersed in a bath at 37 ° C 4h. 用异丙醇透析凝胶以去除未结合的视黄酸,然后在无菌条件下用PBS缓冲液透析。 Dialysis gel with isopropanol to remove unbound retinoic acid, and then dialyzed with PBS buffer under sterile conditions. 将凝胶包装到无菌注射器中并于4°C下储存。 The gel was packaged into a sterile syringe and stored at 4 ° C.

[0185] 实施例15 [0185] Example 15

[0186] 含羟基官能团的添加剂与HA水凝胶的结合。 [0186] Additives containing hydroxyl functional groups bound to the HA hydrogel.

[0187] 添加剂,例如视黄醇(AKA,维甲酸)、过氧化氢酶、二甲氨基乙醇和g-生育酚含有羟基官能团(-0H)。 [0187] Additives, such as retinol (the AKA, retinoic acid), catalase, dimethylaminoethanol and g- tocopherol comprises hydroxy functional groups (-0H). 使用EDC化学经酯化作用,使这些添加剂与HA水凝胶结合。 By esterification using EDC chemistry, these additives are combined with the HA hydrogels. 以下描述了结合的典型实例: The following describes a typical example of binding:

[0188] 使200mg的HMW HA在60cc注射器中IOml的2- (N-吗啉)乙磺酸(MES)缓冲液(pH 4.8)中水合。 [0188] so that the HMW HA 200mg IOml in 60cc syringe 2- (N- morpholino) ethanesulfonic acid (MES) buffer (pH 4.8) hydrated. 在另一个注射器中,将200mg的视黄醇溶于5ml的水-丙酮混合物(水/丙酮体积比1: 3)。 In another syringe, the retinoic 200mg dissolved in 5ml of water - acetone mixture (water / acetone volume ratio of 1: 3). 通过注射器连接器混合以上两个注射器约20次。 Mixing about 20 times more than the two syringes via a syringe connector. 然后将197.7mg的EDC和149mg的NHS分别溶于单独注射器中的6ml水中。 Then 149mg of EDC and NHS 197.7mg were dissolved in 6ml water in separate syringes. 装有EDC和NHS的注射器与装有HA和视黄醇的注射器连接,以通过在2个注射器之间来回传递使反应物混合至少20次。 With EDC and NHS syringe connected to the syringe containing the HA and retinol, through the back and forth between the two syringes pass the reaction mixture was at least 20 times. 将混合物储存在一个注射器中并且浸入37°C浴中4h。 The mixture was stored in a syringe and immersed in a bath at 37 ° C 4h. 用异丙醇透析凝胶以去除未结合的视黄醇,然后在无菌条件下用PBS缓冲液透析。 Dialysis gel with isopropanol to remove unbound retinol, and then dialyzed with PBS buffer under sterile conditions. 将凝胶包装到无菌注射器中并于4°C下储存。 The gel was packaged into a sterile syringe and stored at 4 ° C.

[0189] 实施例16 [0189] Example 16

[0190] 含羟基官能团的添加剂与HA水凝胶通过后改性的结合。 [0190] Modified binding with the HA hydrogels by additive containing hydroxyl functional groups.

[0191] 这是一个两步过程。 [0191] This is a two-step process.

[0192] 步骤1 :用EDC/NHS处理交联HA凝胶,例如基于HA的商用真皮填充剂(例如, JUVEDERM气Allergan、Irvine CA或;Restylane^Medicis Aesthetics,Inc.)以活化HA的羧基。 [0192] Step 1: using EDC / NHS cross-linked HA gel process, for example, commercial HA-based dermal fillers (e.g., gas Juvederm Allergan, Irvine CA, or; Restylane ^ Medicis Aesthetics, Inc.) To activate the carboxyl group of HA.

[0193] 步骤2:用含羟基的添加剂处理活化HA水凝胶。 [0193] Step 2: treated with additives activated hydroxyl group-containing HA hydrogels. 含羟基的添加剂为视黄醇、过氧化氢酶、二甲氨基乙醇和g-生育酚羟基官能团(-0H)。 Additives containing hydroxyl retinol, catalase, dimethylaminoethanol and g- tocopherol hydroxyl functional groups (-0H).

[0194] 添加剂与交联HA凝胶结合的典型实例如下: [0194] Typical examples of the additive combination of crosslinked HA gel as follows:

[0195] 在室温下使2gm的Juvederm凝胶与200gm的EDC和150mg的NHS混合。 [0195] 2gm so Juvederm gel is mixed at room temperature with 150mg of EDC and NHS 200gm of. 然后添加200mg 在3ml丙酮-水混合物中的视黄醇。 200mg acetone was then added at 3ml - retinol water mixture. 使以上混合物在37°C下反应4h。 So that the above reaction mixture for 4h at 37 ° C. 用异丙醇透析凝胶以去除未结合的视黄醇,然后在无菌条件下用PBS缓冲液透析。 Dialysis gel with isopropanol to remove unbound retinol, and then dialyzed with PBS buffer under sterile conditions. 将凝胶包装到无菌注射器中并于4°C下储存。 The gel was packaged into a sterile syringe and stored at 4 ° C.

[0196] 实施例17 [0196] Example 17

[0197] 生长因子、肽或弹性蛋白与HA水凝胶的结合 Binding [0197] growth factor, a peptide or elastin HA hydrogel with

[0198] 含有胺官能团的添加剂,例如表皮生长因子(EGF)、转化生长因子(TGF)和肽,可与HA结合以形成有益真皮填充剂。 [0198] Additives containing amine functional groups, such as epidermal growth factor (EGF), transforming growth factor (TGF) and a peptide, may be combined to form advantageous to HA dermal fillers. 这些添加剂通过酰胺化化学与HA结合。 These additives chemically bound by amidation with HA. 以下描述了结合的典型实例: The following describes a typical example of binding:

[0199] 使200.3mg的HMW HA在IOml的MES pH 5.4缓冲液中水合。 [0199] that the HMW HA 200.3mg hydrated in buffer 5.4 in MES pH IOml. 添加20mg在IOOmg MES溶液中的EGF。 Add 20mg EGF in IOOmg MES solution. 向以上混合物,添加197.7mg的EDC和149mg。 To the above mixture was added 197.7mg of EDC and 149mg. 使所得反应混合物在37°C下反应4h。 The resulting reaction mixture for 4h at 37 ° C. 反应完成后,用异丙醇进一步透析凝胶,然后在无菌条件下用PBS缓冲液透析。 After completion of the reaction, the gel is further dialyzed with isopropanol, and then dialyzed with PBS buffer under sterile conditions. 将凝胶包装到无菌注射器中并于4°C下储存。 The gel was packaged into a sterile syringe and stored at 4 ° C.

[0200] 本发明进一步提供了增强移植脂肪组织生活力的方法。 [0200] The present invention further provides a method of enhancing the viability of transplanted adipose tissue. 所述方法可能通常包括向患者邻近移植脂肪组织的皮肤引入组合物的步骤,所述组合物为本文其它地方描述的组合物。 The method may include the step of generally adjacent the skin graft is introduced to the patient the adipose tissue composition, the composition is a composition thereof as described elsewhere herein. 例如,所述组合物可包含透明质酸和与透明质酸共价结合的维生素C衍生物,其中结合度介于约3mo 1 %和约40mo 1 %之间。 For example, the composition may comprise hyaluronic acid and vitamin C derivative is covalently bound to the hyaluronic acid, wherein the degree of binding between about 3mo 1% and about 40mo 1%. 在本发明的其它方面,治疗皮肤的方法包括向皮肤内引入包含脂肪组织、透明质酸和与透明质酸结合的维生素C的组合物的步骤。 In other aspects of the present invention, a method of treating skin comprising introducing into the skin, adipose tissue, comprising the step of binding the composition of hyaluronic acid and hyaluronic acid and vitamin C.

[0201] 实施例18 [0201] Example 18

[0202] 生长因子、肽或弹性蛋白与HA水凝胶的结合 Binding [0202] growth factor, a peptide or elastin HA hydrogel with

[0203] 为评估维生素C及其衍生物对人脂肪组织来源的干细胞(hASC)的促有丝分裂作用,hASC在组织培养塑料上,于补充或未补充呈游离形式的维生素C (抗坏血酸)或其衍生物(维生食物或AA2G)的完全MesenPro培养基(Invitrogen,Carlsbad,CA)中培养4天。 [0203] To evaluate the pro-vitamin C and derivatives in human adipose tissue-derived stem cells (hASC) of mitosis, hASC in plastic tissue culture, in free form supplemented or Vitamin C (ascorbic acid) or a derivative thereof was (or Vitagen of AA2G) MesenPro complete medium (Invitrogen, Carlsbad, CA) for 4 days. 通过如生产商(ATCC,Manassas,VA)所描述的MTT测定法估计增殖。 The MTT assay by the manufacturer (ATCC, Manassas, VA) as described estimated proliferation. 4天后,发现抗坏血酸0.25、0.5 和ImM的浓度使增殖(按通过脱氢酶转化为紫色甲膳的黄色四唑MTT的量测量)(紫色甲臑被洗涤剂溶解)分别比缺乏抗坏血酸的对照提高60%、80%和96%。 After 4 days, the concentration of ascorbic acid was found that the proliferation of 0.25, 0.5 and ImM (by conversion to purple formazan by dehydrogenase measured amount of yellow tetrazolium MTT-board) (Ru purple formazan dissolved detergent) were higher than controls lacking ascorbic acid 60%, 80% and 96%. 使用相同浓度的AA2G分别获得超过对照70%、60%和50%的增殖提高。 AA2G same concentration over the control respectively 70%, 60% and 50% increase proliferation. 用维生食物获得相似结果,分别显示比对照增加70 %、60 %和30 %。 Similar results were obtained with Vitagen, they show a 70% increase compared with the control, 60% and 30%. 总之,维生素C及其衍生物、AA2G和维生食物在含生长因子的培养基存在下提高了细胞培养中的MSC增殖。 In summary, vitamin C and derivatives thereof, and of AA2G Vitagen MSC increased presence of proliferating cells cultured in a medium containing growth factors.

[0204] 具有结合维生素C的交联HA凝胶 [0204] Vitamin C in combination with a crosslinked HA gel

[0205] 在下面实施例19和20中描述了具有结合维生素C的基于交联HA的凝胶和使用1,4_ 丁二醇二缩水甘油醚(BDDE)作为交联剂的制剂,根据本发明的某些实施方案,其表现出廷德尔效应减小和其它优点。 [0205] In the following Example 19 and 20 described in conjunction with vitamin C-based and cross-linked HA gel using 1,4_ butanediol diglycidyl ether (BDDE) formulation as a crosslinking agent, according to the present invention certain embodiments which exhibit reduced Tyndall effect and other advantages. 在实施例19中,维生素C衍生物是抗坏血酸2-葡萄糖苷(AA2G)而在实施例20中,维生素C衍生物是抗坏血酸3-氨丙基磷酸酯(维生食物)。 In Example 19, the vitamin C derivative is ascorbyl 2-glucoside (of AA2G) In Example 20, the vitamin C derivative is 3-aminopropyl ascorbyl phosphate (vitamin food). 这些凝胶具有最佳流变性、优良的可注射性和高HA浓度(25mg/g)。 These gels have the best rheological properties, excellent injectability and high HA concentrations (25mg / g). 虽然不希望受任何特定操作理论约束,但是本发明人已经发现在AA2G或维生食物存在下使HA与BDDE交联大大改变了凝胶的性质,相对于和BDDE交联的商用HA凝胶,凝胶具有高交联密度、高HA浓度、低粘性和低挤压力。 While not wishing to be bound by any particular theory, the present inventors have found that the presence of HA and BDDE crosslinked gel properties significantly changed with respect to the commercial and BDDE crosslinked HA gel or AA2G Vitagen, gels having a high crosslink density, high HA concentration, low viscosity and a low pressing force. 因为交联期间存在AA2G或维生食物,所以目前形成的凝胶具有单独或经由BDDE,作为侧基和桥接HA链的交联剂与HA链偶联的这些抗坏血酸衍生物。 Since the presence or AA2G Vitagen during the crosslinking, the gel formed with a separate current through BDDE, such as ascorbic acid derivative or a side chain groups and the crosslinker bridged HA HA chains coupled. 凝胶的显微结构已经改变,导致凝胶即使通过细至30G的针,挤压力也非常低。 Gel microstructure has changed, leading to gelation even through a fine needle to 30G, the extrusion pressure is very low. 而且,凝胶具有约3mol %至约IOmol %或高达约15mol %的与HA结合的维生素C。 Moreover, the gel having about 3mol% to about IOmol% up to about 15mol%, or bound to the HA vitamin C. 在注射凝胶时,它们通过内源酶,例如来自成纤维细胞的α-葡萄糖苷酶或磷酸酶释放活性维生素C。 When injectable gel, which by endogenous enzymes, such as fibroblasts derived from α- glucosidase or phosphatase release the active vitamin C. 活性维生素C可引发皮肤胶原蛋白形成并且可起自由基清除剂的作用抑制凝胶降解。 Active vitamin C can cause the formation of collagen and skin can act as a radical scavenger to inhibit the degradation of the gel.

[0206] 实施例19 [0206] Example 19

[0207] 廷德尔效应减小的HA/AA2G凝胶的配制 Formulation [0207] Tyndall effect reduced HA / AA2G gel

[0208] 在添加1764. Omg的5wt %NaOH溶液后,使注射器中400 . Img的LMW HA和402.3mg AA2G的混合物水合60min。 [0208] After the addition of 1764. Omg 5wt% NaOH solution, so that the syringe 400. Img LMW HA and the mixture of hydrous 60min 402.3mg AA2G. 在单独小瓶中添加800.8mg的AA2G,接着添加1401. Img的9. lwt% NaOH溶液和252.6mg的BDDE。 AA2G 800.8mg added in a separate vial, followed by addition of 9. lwt% NaOH solution 1401. Img and 252.6mg of BDDE. 使所得溶液(pH> 12)在50°C水浴中反应〜20min,之后将其转移到水合HA中。 The resulting solution (pH> 12) 50 ° C and the reaction ~20min water bath, then transferred to the hydrated HA. 添加之后,通过将其在两个注射器之间来回传递使混合物混合〜20次。 After the addition, by passing it back and forth between the two syringes mixed and the mixture was ~ 20 times. 然后将糊剂转移到小瓶中,之后将其置于50°C水浴中〜2.5h。 The paste is then transferred to a vial, then placed in a water bath at 50 ° C ~2.5h. 交联后,添加含197.Omg的12M HCl 和9.18gl0 X PBS (pH 7.4)的溶液以中和碱,并且使凝胶在定轨振荡器上膨胀72h。 After crosslinking, adding a solution of 12M HCl and containing 197.Omg 9.18gl0 X PBS (pH 7.4) to neutralize the base, and the Gel Expansion 72h on an orbital shaker. 迫使其通过〜60μπι孔径的网筛分凝胶。 Forced through a sieve mesh aperture ~60μπι gel. 通过将其在两个注射器之间来回传递混合筛过的凝胶〜20 次,之后将其转移到纤维素酯透析袋(MffCO〜20kDa)中并且用PBS (pH 7.4)缓冲液透析5天, 每天更换缓冲液两次。 Mixed by passing it through a sieve gel ~ 20 back and forth between the two syringes times, then transferred to a cellulose ester dialysis bag (MffCO~20kDa) and treated with PBS (pH 7.4) buffer, dialyzed for 5 days buffer exchanged twice a day. 透析后,将凝胶分配到Iml COC注射器中,在5000RPM下离心5min以去除气泡,并且用湿蒸汽灭菌。 After dialysis, the gel was distributed to Iml COC syringe, centrifuged 5min at 5000RPM to remove air bubbles, and the wet steam sterilization. 该凝胶具有25mg/g的HA最终浓度,如实施例2中所述计算的约10111〇1%的4426 111〇1%和约80?3的6'。 The gel has a final concentration of HA 25mg / g, as in Example 2. The calculated 10111〇1 about 4426% and about 80% 111〇1? 63 & apos. 以相似方式制备其它凝胶,6'值为约60?3至约80?3。 Other gel was prepared in a similar manner, 6 'value of about 60? To about 80 3? 3.

[0209] 实施例19A [0209] Example 19A

[0210] 廷德尔效应减小的具有利多卡因的HA/AA2G凝胶的配制 Formulation [0210] Tyndall effect reduced HA / AA2G gel with lidocaine

[0211] 向实施例19的凝胶添加一定量的利多卡因以生成具有利多卡因的HA/AA2G凝胶, 其具有〇. 3 %ww利多卡因。 HA / AA2G gel [0211] added to the gel a quantity of lidocaine of Example 19 to generate a lidocaine, having a square. 3% ww lidocaine. 通过将利多卡因HCl溶于PBS缓冲液(pH〜7.4)中制备利多卡因溶液。 Lidocaine solution prepared by adding lidocaine HCl was dissolved in PBS buffer (pH~7.4). 透析之后但是在灭菌之前,向实施例19中的凝胶添加等分试样的利多卡因溶液。 But before sterilization, lidocaine solution aliquot is added to the gel in Example 19 after the dialysis. 然后完全混合凝胶以获得具有0.3 % ww利多卡因浓度的均匀混合物。 The gel is then thoroughly mixed to obtain a homogeneous mixture having a concentration of 0.3% ww lidocaine.

[0212] 实施例20 [0212] Example 20

[0213] 廷德尔效应减小的HA/维生食物凝胶的配制 Formulation [0213] Tyndall effect reduced HA / gel Vitagen

[0214] 使401.〇11^的11^似在注射器中2355.〇11^的1¥七%似0!1溶液中水合〜45111丨11。 [0214] ^ 11 ^ make 401.〇11 like in the syringe 2355.〇11 ^ ¥ seven percent of the likelihood of 0 1! 1 was hydrated ~45111 Shu 11. 将303.8mg BDDE添加到水合HA中并且通过注射器间混合,混合10次。 303.8mg BDDE added to the hydrated HA and by inter-mixing syringe and mixed 10 times. 使混合物在50 °C水浴中预先反应15min。 The mixture was pre-reacted 15min 50 ° C and water bath. 将800. Img的维生食物单独溶于950.6mg的15wt%Na0H中,接着溶于510. IMilli-Q水中。 800. Img of the individual Vitagen dissolved in 15wt% Na0H 950.6mg, then dissolved 510. IMilli-Q water. 使用注射器间混合,使维生食物溶液与预热的水合HA/BDDE混合物来回混合30次。 Using the inter-syringe mixing, hydrated HA Vitagen solution preheated / BDDE mixture is mixed back and forth 30 times. 将混合物放回50°C水浴中并且再继续反应2h,之后将含有148. Img的12M HCl和8523. Img的10 X PBS (pH7.4)的溶液添加到交联剂中。 The mixture was returned to 50 ° C water bath and the reaction was continued for 2h, after containing the 12M HCl and 148. Img 8523. Img of 10 X PBS (pH7.4) was added to the crosslinking agent. 添加HCl-I3BS溶液以中和凝胶并使其膨胀。 HCl-I3BS solution was added to neutralize the gel and allowed to swell. 使凝胶中和并且在定轨振荡器上膨胀48h以上。 The gel was neutralized and expanded for 48h on an orbital shaker. 通过〜60μπι筛子筛分凝胶并且通过在2个注射器之间来回传递混合〜20次。 ~60μπι sieved through sieve gel and transferred back and forth between two syringes through a mixing ~ 20 times. 将凝胶放入20,000 MW⑶CE透析袋中并且在PBS (pH7.4)缓冲液中透析。 The gel was placed in a dialysis bag and dialyzed 20,000 MW⑶CE in PBS (pH7.4) buffer. 透析进行〜197h,频繁更换PBS缓冲液。 Dialysis ~197h, frequent change of PBS buffer. 透析后,将凝胶转移到Iml COC注射器中,在5000RPM下离心5min并用湿蒸汽灭菌。 After dialysis, the gel was transferred to Iml COC syringe, centrifuged 5min at 5000RPM with a moist steam sterilization. 凝胶的HA最终浓度为24mg/g。 The final concentration of HA gel is 24mg / g.

[0215] 经1-乙基-3_[3-二甲基氨丙基]碳二亚胺盐酸盐(EDC)化学交联的HA凝胶 [0215] 1-Ethyl--3_ by [3-dimethylaminopropyl] carbodiimide hydrochloride (EDC) chemically cross-linked HA gel

[0216] 在下面实施例21和22中描述了根据本发明的某些实施方案,表现出廷德尔效应减小和其它优点的基于交联HA的凝胶的制备。 [0216] Examples 21 and 22 described in the following embodiment according to certain embodiments of the present invention, prepared exhibit reduced Tyndall effect and other advantages of the crosslinked HA-based gel. 在实施例21中,使用为己二胺(HMDA)的交联剂而在实施例20中使用为3-[3- (3-氨丙基)-2,2-双(3-氨基-丙氧基甲基)-丙氧基]-丙胺(4 臂胺-4AA)的交联剂,经EDC化学制备凝胶。 Crosslinking agent used in Example 21, the use of hexamethylene diamine (of HMDA) in Example 20 3- [3- (3-aminopropyl) -2,2-bis (3-amino - propionic yloxymethyl) - propoxy] - propylamine (amine 4 arm -4AA) crosslinking agent, prepared by gel EDC chemistry. 在温和条件下,例如室温和例如pH 5.4下进行交联。 Under mild conditions, for example at room temperature and crosslink e.g. pH 5.4. 可调整反应条件以制备具有最佳凝胶性质、优良可注射性和高HA最终浓度(〜24mg/g) 的高度网状凝胶。 The reaction conditions are adjusted to produce a gel having the best properties, excellent injectability and high final concentration of HA (~24mg / g) the height of the mesh gel. 发明人已经发现,在非常低的水合或反应浓度下,用适量HMDA或4AA,连同偶联剂,即1-乙基-3- [3-二甲基氨丙基]碳二亚胺盐酸盐(EDC)和N-羟基琥珀酰亚胺(NHS) 或磺酰基-NHS (磺基-NHS)使HA交联可能有利。 The inventors have found, at very low hydration reaction or concentration, or with the amount of HMDA 4AA, together with the coupling agent, i.e., 1-ethyl-3- [3- dimethylaminopropyl] carbodiimide hydrochloride salt (EDC) and N- hydroxysuccinimide (NHS) or sulfonyl group -NHS (sulfo-NHS) may be advantageous that the crosslinked HA. 凝胶的交联点将相互远离,因此高度交联的材料具有高阻尼力。 Crosslinked gel point away from each other, and therefore highly crosslinked material having a high damping force. 相反,HA与BDDE在如此低的水合或反应浓度下的交联可能由于交联剂相对无效而不可行。 Conversely, HA and BDDE crosslinking at such a low concentration of the hydration reaction or a crosslinking agent may be relatively ineffective because not feasible.

[0217] 实施例21 [0217] Example 21

[0218] 廷德尔效应减小的HA/HMDA凝胶的配制 Formulation [0218] Tyndall effect reduced HA / HMDA gel

[0219] 将20.Og的IOOmM MES缓冲液(pH 5.2)添加到装有1000.Omg的LMff HA的注射器中。 [0219] 20.Og of the IOOmM MES buffer (pH 5.2) was added to the LMff HA 1000.Omg containing syringes. 通过将260.9mg HMDA.HCl溶于2010.5mg的IOOmM MES缓冲液(pH 5.2)中,并且添加2μ1的IM NaOH使pH达到5.2制备HMDA溶液。 2010.5mg 260.9mg HMDA.HCl dissolved by the IOOmM MES buffer (pH 5.2), and IM NaOH was added 2μ1 of HMDA solution was prepared to bring the pH reached 5.2. 通过将254.2mg的EDC溶于1188.4mgl00mM MES缓冲液(pH 5.2)中制备EDC溶液,并且于单独小瓶中,将44.3mg的NHS溶于1341.8mg的IOOmM MES缓冲液(pH 5.2)中。 By 254.2mg of EDC EDC dissolved 1188.4mgl00mM MES buffer solution prepared in (pH 5.2), and in a separate vial, 44.3mg of NHS were dissolved in 1341.8mg IOOmM MES buffer (pH 5.2) in. HA完全水合后,〜Ih,向水合HA中添加790μ1的HMDA溶液。 After fully hydrated HA, ~Ih, HMDA solution was added to the hydrated HA 790μ1. 通过注射器间混合匀化混合物10次。 By inter-mixing syringe The mixture was homogenized 10 times. 然后将490μ1 EDC和490μ1 NHS溶液添加到匀化糊剂中并且通过注射器间混合再混合10次。 Then 490μ1 EDC and 490μ1 NHS solution was added to the homogenized paste and by inter-mixing syringe further mixing 10 times. 然后将混合物转移到小瓶中并且在添加17.9ml的IX PBS缓冲液(pH 7.4)之前,在室温下交联5h。 Before The mixture was then transferred to a vial and adding 17.9ml of IX PBS buffer (pH 7.4), crosslinked 5h at room temperature. 在迫使其通过60μπι孔径的网之前,使凝胶在滚筒上膨胀3天。 Prior to forcing it through a mesh aperture 60μπι the gel expands in the drum 3 days. 将筛分的凝胶放在纤维素酯膜透析管MffCO 20KDa中并且用I XPBS透析4天,每天更换缓冲液两次。 The sieved cellulose ester film placed on the gel in a dialysis tube with I XPBS MffCO 20KDa and dialyzed for 4 days changing the buffer twice daily. 将凝胶分配在Iml COC注射器中,在5000RPM下离心5min,并用湿蒸汽灭菌。 The gel was partitioned Iml COC syringe, centrifuged at 5000RPM 5min, and wet steam sterilization. 凝胶的HA最终浓度为25mg/g。 The final concentration of HA gel is 25mg / g.

[0220] 实施例22 [0220] Example 22

[0221] 廷德尔效应减小的HA/4AA凝胶的配制 [0221] Tyndall effect reduced HA / 4AA gel formulation

[0222] 将32.55g的IOOmM MES缓冲液(pH 5.2)添加到装有1000.4mg的LMW HA的注射器中。 [0222] 32.55g of the IOOmM MES buffer (pH 5.2) was added to the syringe containing the LMW HA 1000.4mg. 通过将256.3mg 4AA溶于1039.8mg的IOOmM MES缓冲液(pH 5.2)中,并且添加380μ1的6M HCl使pH达到5.2制备4ΑΑ溶液。 1039.8mg 256.3mg 4AA dissolved by the IOOmM MES buffer (pH 5.2), and added 380μ1 6M HCl to reach a pH of 5.2 was prepared 4ΑΑ. 通过将251.2mg的EDC溶于1013.8mg IOOmM MES缓冲液(pH 5.2)中制备EDC溶液,并且于单独小瓶中,将74.7mg的NHS溶于2020. Omg的IOOmM MES缓冲液(pH 5.2)中。 By 251.2mg of EDC EDC solution prepared in (pH 5.2) was dissolved 1013.8mg IOOmM MES buffer, and in a separate vial, 74.7mg of NHS were dissolved in 2020. Omg IOOmM MES buffer (pH 5.2) in. HA完全水合后,〜Ih,向水合HA中添加260μ1的4AA溶液。 After fully hydrated HA, ~Ih, 4AA solution was added to the hydrated HA 260μ1. 通过注射器间混合匀化混合物10次。 By inter-mixing syringe The mixture was homogenized 10 times. 然后将277μ1 EDC和273μ1 NHS溶液添加到匀化糊剂中并且通过注射器间混合再混合10次。 Then 277μ1 EDC and 273μ1 NHS solution was added to the homogenized paste and by inter-mixing syringe further mixing 10 times. 然后将混合物转移到小瓶中并且在添加6.4ml的10 X PBS缓冲液(pH 7.4)之前,在室温下交联5h。 The mixture was then transferred to a vial and 10 X PBS before 6.4ml of buffer (pH 7.4) is added, crosslinking 5h at room temperature. 在迫使其通过60μπι孔径的网之前,使凝胶在滚筒上膨胀3天。 Prior to forcing it through a mesh aperture 60μπι the gel expands in the drum 3 days. 将筛分的凝胶放在纤维素酯膜透析管MffCO 20KDa中并且用I XPBS透析4天,每天更换缓冲液两次。 The sieved cellulose ester film placed on the gel in a dialysis tube with I XPBS MffCO 20KDa and dialyzed for 4 days changing the buffer twice daily. 将凝胶分配在Iml COC注射器中,在5000RPM下离心5min,并用湿蒸汽灭菌。 The gel was partitioned Iml COC syringe, centrifuged at 5000RPM 5min, and wet steam sterilization. 凝胶的HA最终浓度为23mg/g。 The final concentration of HA gel is 23mg / g.

[0223] 实施例23 [0223] Example 23

[0224] 实施例19-22的凝胶流变性的测定 The gel rheology Example 19-22 [0224] Embodiment

[0225] 使用振荡平行板流变仪,Anton Paar Physica MCR 301,测量凝胶的流变性。 [0225] Using the oscillating parallel plate rheometer, Anton Paar Physica MCR 301, the measurement of the gel rheology. 在间隙高度为Imm时使用25mm的板直径。 Using 25mm diameter plates at a gap height of Imm. 在25 °C恒温下进行测量。 Measured at 25 ° C for constant temperature. 每次测量由2 %恒定应变和频率对数增长下I-IOHz的频率扫描组成,接着是随应变对数增长,5Hz恒定频率下1-300 %的应变扫描。 Each measurement a constant strain of 2% and a frequency of the frequency sweep logarithmic growth composition of the I-IOHz, followed with strain logarithmic growth, 5Hz constant frequency scanning 1-300% strain. 在1 %应变下由应变扫描获得储能模量(G')和粘性模量(G”)。 Obtained from a strain sweep the storage modulus (G ') at 1% strain and a viscous modulus (G ").

[0226] 由实施例19-22获得的凝胶的储能模量和粘性模量 [0226] The storage modulus and the viscous modulus of Examples 19-22 obtained by the gel

Figure CN108379112AD00331

[0228] 实施例24 [0228] Example 24

[0229] 实施例19-22的凝胶的挤压力测量 Examples 19-22 Gel pressing force of the [0229] embodiment of the measurement

[0230] 使用Instron 5564和Bluehill 2软件测量将凝胶挤出30G针所需的力。 [0230] using the Instron 5564 and measured Bluehill 2 software extruding the gel force required 30G needle. 将凝胶从Iml COC注射器挤出30G1/2 TSK针。 The gel was extruded 30G1 / 2 TSK Iml COC needle from the syringe. 按100mm/min的速度推动活塞11.35min,并记录挤压力。 Press speed of 100mm / min to push the piston 11.35min, and record the pressing force.

[0231] 由实施例19-22获得的凝胶的挤压力 [0231] Examples 19-22 obtained by the pressing force of the gel

Figure CN108379112AD00332

[0233] 实施例25 [0233] Example 25

[0234] 实施例19-22的凝胶的生物相容性试验 [0234] Examples 19-22 gel biocompatibility tests embodiment

[0235] 在史-道二氏大鼠(Sprague Dawley rat)的背面皮内植入50μ1弹丸注射的凝胶。 [0235] In the history - a gel implant 50μ1 bolus injection within the skin back surface two channels Dawley rats (Sprague Dawley rat) is. 在1周时取出植入物并用苏木精和伊红(Η&Ε)染色和是单核炎性细胞标志物的⑶68染色,通过组织学分析植入物。 1 taken at the implant periphery and stained with hematoxylin and eosin (Η & amp; Ε) ⑶68 staining and staining mononuclear inflammatory cells markers, by histological analysis of the implant. 基于染色程度为⑶68的3张20 X图像评分0-4。 Based on the degree of staining ⑶68 of three 20 X image score 0-4. 然后平均这些值以给出样品得分。 These values ​​were then averaged to give a score of the sample. 每种凝胶分析4个样品。 Four samples of each gel analysis.

[0236] 实施例19-22的平均⑶68得分 The average ⑶68 [0236] Examples 19-22 score

Figure CN108379112AD00341

[0239] 实施例26 [0239] Example 26

[0240] 实施例19-22的凝胶的细胞毒性试验,ISO 10993-5。 [0240] Examples 19-22 toxicity test gel cells embodiment, ISO 10993-5.

[0241] 根据ISO 10993-5的琼脂糖覆盖法,通过NAMSA进行凝胶的体外细胞毒性试验:《医疗器械生物学评价》-第5部分:体外细胞毒性试验。 [0241] The ISO 10993-5 agarose overlay method, the in vitro cytotoxicity test gel by NAMSA: "Biological Evaluation of Medical Devices" - Part 5: in vitro cytotoxicity assays. 向三个复孔添加0.1ml放在滤盘上的试验样品,以及0.9%NaCl溶液、作为阴性对照的Icm长的高密度聚乙烯和作为阳性对照的IX Icm2乳胶部分。 0.1ml was added to triplicate wells in a test sample on the filter plate, and a 0.9% NaCl solution, as long Icm high density polyethylene and IX Icm2 latex portion of the negative control as a positive control. 将每一个放在直接覆盖单层L929小鼠成纤维细胞的琼脂糖表面。 In each of a single layer directly covers the surface of the agarose into mouse L929 fibroblasts. 37°C下在5%C02中孵育24h后,从宏观和微观上检查培养物的任何异常细胞形态和细胞裂解。 After 24h at 37 ° C for the 5% C02, check culturing any abnormal cellular morphology and cell lysates from macroscopically and microscopically. 基于靠近样品的裂解区,为试样评分0-4。 Based on the cracking zone close to the sample, a sample score 0-4. 因为试样样品未显示出引起任何细胞裂解或毒性的迹象,所以来自实施例1、3和4的试验材料得分为0。 Because the sample to cause the sample did not show any signs of toxicity or cell lysis, and so from Example 3 Test material 4 0 score.

[0242] 廷德尔效应的定量分析 Quantitative analysis [0242] Tyndall effect

[0243] 为了进一步支持目视观测和进行HA填充剂的比较优势分析,认为必须进行廷德尔效应的定量分析。 [0243] In order to further support the visual observations and carry out comparative advantage analysis of HA fillers, considered necessary for quantitative analysis Tyndall effect. 同样在文献中没有对真皮填充剂有特异性的廷德尔效应的定量技术。 Also there is no Tyndall effect specific quantification of dermal fillers in the literature. 然而,基于对光散射及光和皮肤相互作用的现有科学理解,采用基于(a)比色法和(b)光谱学的两种不同方法量化皮肤上的廷德尔效应。 However, based on current scientific understanding light scattering and light skin interaction, quantified using Tyndall effect on the skin of two different methods (a) and colorimetry (b) based on spectroscopy. 基于这些技术,定义了3种不同的定量参数(如下所示)来测量体内廷德尔效应。 Based on these technologies, we define three different quantitative parameters (see below) to measure body Neitingdeer effect.

[0244] a)廷德尔效应目测得分:标度范围为1-5,增量为0.5。 [0244] a) Tyndall effect visual score: 1-5 scale ranges, 0.5 increments. 对肤色正常且没有蓝色变色的注射部分给予得分1。 Score given one pair of normal skin and no portion of the blue discoloration injection. 对厚且明显的蓝色变色(通常与Restylane或Juv6derm Ultra Plus 相关)给予最高得分5。 Of thick and clear blue color (typically associated with Restylane or Juv6derm Ultra Plus) 5 given the highest score. 在为试样盲评分之前,为3名独立观察者培训标度。 Before score for the blind sample, three independent observers for the training scale.

[0245] b)皮肤颜色的蓝色组分_“b” :使用色度计(CM2600D,Konica Minolta,NJ)量化从注射了不同填充剂的皮肤部位出射的光的蓝色组分。 [0245] blue component b) skin color _ "b": quantization of light from the skin site injected with different fillers emitted blue component using a colorimeter (CM2600D, Konica Minolta, NJ). 这通过使用Lab色标的“b”组分实现。 This "b" components is achieved by using the Lab color scale.

[0246] c)从皮肤出射的“蓝光%”:使用便携式分光光度计(CM2600D,Konica Minolta, NJ)量化总可见光范围内从皮肤出射的蓝光%。 [0246] c) emitted from the skin "% blue": using a portable spectrophotometer (CM2600D, Konica Minolta, NJ) quantization blue light emitted from the skin% of the total visible range. 这可通过求400-490nm可见光谱下的面积的积分并且用光谱(400-700nm)下的总面积将其标准化而实现。 This is accomplished by integrating the area under the 400-490nm and the visible spectrum with total area under the spectrum (400-700 nm) to achieve standardization.

[0247] 实施例27 [0247] Example 27

[0248] 凝胶的廷德尔评估 [0248] Evaluation gel Tyndall

[0249] 使用直线注射法,通过27G1/2 TSK针向2月龄无毛大鼠的大腿皮内注射凝胶。 [0249] A straight-line injection method, the 2-month-old hairless rats by 27G1 / 2 TSK gel intradermal injection needle inside the thigh. 浅表植入凝胶以模拟临床细纹程序。 Superficial gel implant analog lines clinical procedures. 在凝胶植入后进行廷德尔试验48h。 Tyndall test performed 48h after gel implantation. 进行廷德尔试验后,使动物安乐死以提高由于缺乏血红蛋白,廷德尔效应的对比度。 After Tyndall test, the animals were euthanized due to a lack of increase of hemoglobin, the contrast Tyndall effect.

[0250] 图12中示出了植入2天后,来自实施例19和21的凝胶的图像。 [0250] FIG. 12 shows the implant of 2 days, from the image of the gel in Example 19 and 21 embodiment. 还示出了商用Juv6 derm Refine和Restylane Touch的图像以便比较。 The image also shows for comparison a commercial Juv6 derm Refine and Restylane Touch of. 在商用Juv6derm Refine和Restylane Touch的图像中蓝色线(廷德尔效应)清晰可见。 In commercial Juv6derm Refine and Restylane Touch the blue line in the image (Tyndall effect) visible. 来自实施例19、19A (未示出)和21的凝胶未表现出廷德尔效应。 From Example 19, 19A (not shown) and the gel 21 does not exhibit Tyndall effect.

[0251] 使用增量为0.5的目测得分1-5,为注射部位评分。 [0251] 0.5 using increments of visual score 1-5 score injection site. 得分为1的注射部位未显示出皮肤变色,而得分为5的注射部位显示出皮肤严重的蓝色变色。 A score of 1 injection sites showed no discoloration of the skin, a score of 5 shows the injection site blue serious skin discoloration. 还借助于色度计(CM2600D, Koni ca Mino I ta,NJ)对注射部位进行了光谱分析。 Also by means of a colorimeter (CM2600D, Koni ca Mino I ta, NJ) to the injection site were spectrometry. 皮肤颜色的蓝色组分“b”和从皮肤出射的蓝光% (400_700nm)独立测量。 Blue skin color component "b" and (400_700nm) independent measurements% of blue light emitted from the skin. 图13和14示出了目测廷德尔得分和出射蓝光%。 13 and 14 illustrate a visual score and exit blue Tyndall%. 来自实施例19和21的凝胶未表现出廷德尔效应并且具有较低的目测廷德尔得分和出射蓝光值%。 Gels from Examples 19 and 21 embodiment does not exhibit a Tyndall effect and having a low score and visual exit blue Tyndall% value. 对于Juv6derm Ref ine和Re sty lane Touch而言,廷德尔得分和出射蓝光值%较高。 For Juv6derm Ref ine and Re sty lane Touch, the exit points and blue Tyndall% of the higher value. Be Iotero Soft未显示出廷德尔效应并且值比得上实施例19和21的凝胶。 Be Iotero Soft showed no Tyndall effect and a value comparable to the gel of Examples 19 and 21, respectively. 见图13和14。 Figures 13 and 14.

[0252] 实施例28 [0252] Example 28

[0253] 通过组织学对凝胶的体内持续时间评估 [0253] Studies in vivo evaluation of the duration of the gel through the tissue

[0254] 在史-道二氏大鼠的背面皮内植入50μ1弹丸注射的本发明的凝胶和商用凝胶。 [0254] In the history - and commercial gel gel implants of the present invention 50μ1 bolus injection in the back of the skin-Jakob tract of rats. 在1 周时取出植入物并用苏木精和伊红(Η&Ε)染色通过组织学分析。 The implant taken at 1 week and stained with hematoxylin and eosin (Η & amp; Ε) by staining for histological analysis. 就在注射部分取切片。 Sections were taken on the injection portion. 从每个组织样品切两个切片并且使用缝合指示器缝合Η&Ε染色切片。 Cut two sections from each tissue sample and the use of stitching sewn indicator Η & amp; Ε stained sections. 然后将样品分组并且根据剩余材料的量评分如下:没有(〇%)、低(25%)、中(50%)和高(100%)。 The samples were then grouped according to the amount and the scores of the remaining material follows: No (square%), low (25%) (50%) and high (100%). 见图15。 See Figure 15.

[0255] 实施例28Α [0255] Example 28Α

[0256] 通过MRI对凝胶的体内持续时间评估 [0256] Evaluation of the duration of the gel in vivo by MRI

[0257] 使用磁共振成像(MRI)研究评估在雌性史-道二氏大鼠中皮内注射后,40周内本发明凝胶和商用凝胶的体积和表面积随时间的变化。 [0257] using a magnetic resonance imaging (MRI) study evaluated in female history - Jakob channel rats after intradermal injection, and the volume of the gel 40 weeks of the present invention and a commercial gel surface area over time. 按每种植入物150μ1靶体积注射凝胶。 Per target volume of implant 150μ1 injectable gels. 使植入物位于肩膀稍近尾部的两个对侧部位、稍远离膝盖吻侧的两个对侧部位和中点介于头尾之间的两个对侧部位。 Two opposite sides of the implant site is located near the rear shoulder slightly, slightly away from the two opposite sides of the knee rostral site, and the midpoint between two opposite side portions between the head and tail. 可在7Tesla 70/30 Bruker Biospec MRI扫描仪上进行扫描。 It can be scanned on 7Tesla 70/30 Bruker Biospec MRI scanner. 在植入当天(第〇周)和植入后12、24、40周收集图像。 12,24,40 weeks after implantation and collected images of the day implantation (first billion Week). 下面图16中示出了凝胶绝对体积与时间的图。 Figure 16 below shows the absolute volume of the gel in FIG time. 高持久性凝胶在植入40周时具有高绝对体积。 Persistent high gel having a high absolute volume at 40 weeks implantation.

[0258] 实施例29 [0258] Example 29

[0259] 用于治疗眶周纹的本发明组合物 [0259] for the treatment of periorbital lines of the compositions of the invention

[0260] 40岁的瘦小女人在眶周区出现细小皱纹并且要求真皮填充剂治疗。 [0260] skinny 40-year-old woman appeared fine wrinkles in the periorbital region and requires dermal filler treatment. 使用30G针,医师向她每只眼睛下方和泪沟区中的细纹浅表引入0.6ml根据本发明所述基于HA的凝胶(例如实施例19中描述的凝胶)。 Use 30G needle, her physician and below each eye of the tear lines in shallow trench regions introduced into 0.6ml HA-based gels according to the present invention (e.g. gel described in Example 19). 虽然在浅表引入凝胶,但是未观察到蓝色变色并且患者对结果满意。 Although the introduction of superficial gel, but no blue color was observed and the patient is satisfied with the results.

[0261] 正如所示,本发明的组合物,例如实施例19和21的组合物,廷德尔效应减小或不明显,并且相对于某些基于HA的商用凝胶,例如Juvederm Refine/Surgiderm 18和Belotero Soft,在体内的持续时间长很多。 [0261] As indicated, the compositions of the invention, e.g. Example 19 and 21 compositions, the Tyndall effect is reduced or insignificant embodiment, with respect to certain commercial HA-based gels, e.g. Juvederm Refine / Surgiderm 18 and Belotero Soft, in the body much longer duration. 例如,相对于商用“细纹”制剂Belotero Soft,本发明的实施例19不但具有至少与这种商用凝胶一样有利的廷德尔得分,而且有利地表现出高很多的体内持续时间。 For example, with respect to the commercial "lines" formulation Belotero Soft, embodiments of the present invention not only has at least 19 such as a commercial gel advantageous Tyndall score, and advantageously exhibit a much higher duration in vivo.

[0262] 实施例30 [0262] Example 30

[0263] 用于改善细纹出现的本发明的可注射组合物 [0263] The present invention for improving the appearance of fine lines of the injectable compositions

[0264] 单独和组合的添加剂,例如维生素A、维生素B、维生素C、维生素D、维生素E及其衍生物以便于生成多种大体上光学透明、可注射的基于HA的凝胶的方式与交联透明质酸凝胶结合。 [0264] alone, and combinations of additives, such as vitamin A, vitamin B, vitamin C, vitamin D, vitamin E and its derivatives in order to generate a variety of substantially optically transparent, HA-based gel manner injectable delivery combined with hyaluronic acid gel. 如本文其它地方所定义,按重量计HA组分至少90%,例如大体上完全为低分子量HA, 或约100%低分子量HA。 As defined elsewhere herein, by weight HA component of at least 90%, such as low molecular weight HA is substantially complete, or about 100% of low molecular weight HA. 使用任何适合方式使这些添加剂与HA水凝胶结合。 Using any suitable manner such additives in combination with HA hydrogels. 筛分并加工结合的凝胶以生成可注射、pH中性、粘性组合物,其HA浓度至少约20mg/g,例如约23、约24mg/g、 约25mg/g、高达约30mg/g并且适合通过细规格针注射。 Sieved and processed to generate combined injectable gel, pH-neutral, tacky composition which HA concentration of at least about 20mg / g, for example about 23, to about 24mg / g, from about 25mg / g, up to about 30mg / g and suitable for injection through a narrow gauge needle. 凝胶的G'值至少约50PA、约60Pa、约70Pa、约80Pa,高达或不大于约lOOPa。 Gel G 'value of at least about 50PA, about 60Pa, about 70Pa, about 80Pa, or no more than up to about lOOPa. 包装凝胶并使用高压锅、紫外光或其它适合方式灭菌。 And packaging the gel using an autoclave, UV sterilization or other suitable manner.

[0265] 每种凝胶对浅表注射均有用,例如在不大于约I. Omm的深度注射到患者皱纹,例如眶周区、鼻唇沟区、泪沟区、颈部区或将受益于真皮填充的任何其它面部区域的皮肤中。 [0265] Each of the gel for superficial injections are used, for example, at a depth of no more than about I. Omm wrinkles injected into a patient, e.g. periorbital area, nasolabial area, tear furrows, or neck region would benefit any other facial skin areas of dermal filler. 尽管浅表引入凝胶,但是未观察到由于廷德尔效应的变色并且患者满意结果。 Although the superficial introduction of gel, but the discoloration was not observed due to the Tyndall effect results and patient satisfaction.

[0266] 最后,应理解,虽然已参考各实施方案描述了本说明书的方方面面,但是本领域的技术人员将易于认识到,公开的特定实施例仅仅是说明了本文公开的主题原理。 [0266] Finally, it should be understood that, although the embodiment has been described with reference to various aspects of embodiments of the present specification, those skilled in the art will readily recognize that the disclosed embodiments are merely illustrative of particular embodiments of the principles of the subject matter disclosed herein. 因此,应理解,公开的主题决不限于本文描述的特殊方法、方案和/或试剂等。 Therefore, it should be understood that the disclosed subject matter described herein in no way limited to the particular methods, protocols, and / or reagents. 因而,在不背离本说明书的精神的前提下,本领域的技术人员可根据本文的教导对公开的主题做许多不同修改或变化或替代构型。 Thus, without departing from the spirit of the present specification, the premise of the present skilled in the art may make various modifications or variations or a number of alternative configurations of the disclosed subject matter in accordance with the teachings herein. 在不背离所附权利要求中定义的本发明精神的前提下,可做详细变化。 Premise defined in the appended claims without departing from the spirit of the invention, changes in detail do. 最后,本文使用的术语仅仅是为了描述特定实施方案,而非旨在限制本发明的范围,该范围仅由权利要求定义。 Finally, the terminology used herein is for describing particular embodiments only embodiment, not intended to limit the scope of the invention, which scope is defined solely by the claims. 另外,其意图是以上描述中所含的或附图中示出的所有材料应该仅解释为说明性而非限制性。 Further, it is intended that the above description contains all materials or illustrated in the drawings should be construed as merely illustrative and not restrictive. 因此,本发明不限于精确显示和描述的那些。 Accordingly, the present invention is not limited to those shown and described accurately.

[0267] 本文描述了本发明的某些实施方案,包括发明人已知用于进行本发明的最佳模式。 [0267] Certain embodiments herein described embodiment of the present invention, comprising known to the inventors for carrying out the best mode of the present invention. 当然,描述的这些实施方案的变型对于本领域的普通技术人员在阅读前述描述后将会变得显而易见。 Of course, variations of these embodiments described with respect to those of ordinary skill in the art will become apparent upon reading the foregoing description. 发明人期望技术人员视情况采用此类变型,而且发明人意在使本发明以不同于本文具体描述的方法实施。 The inventors expect skilled artisans to employ such variations, and the inventors of the present invention is intended to a method different from the embodiments specifically described herein. 因此,如适用法律许可,本发明包括所附权利要求中叙述的主题的所有修改和等同形式。 Accordingly, all such modifications permitted by applicable law, according to the present invention include the subject matter recited in the appended claims and equivalents. 而且,除非本文另外指出或另外与上下文明显矛盾,否则本发明涵盖其所有可能变型中的上述元素的任何组合。 Furthermore, unless otherwise indicated herein or otherwise clearly contradicted by context, the present invention encompasses any combination of or all possible variations of the above-described elements.

[0268] 不应将本文公开的本发明的替代元素或实施方案的分组理解为限制。 [0268] Alternatively the packet should not be elements of the invention disclosed herein or embodiments be construed as limiting. 每个组的成员可单独或以与该组其它成员或本文找到的其它元素的任何组合提及和要求保护。 Each group member may be used alone or in any combination with other elements found in other members of the group or mentioned and claimed herein. 预计, 组的一个或多个成员可因方便和/或专利性的原因包括在组中或从中删除。 We expect one or more members of the group may be due to reasons of convenience and / or patentability included in the group or remove. 当任何此类包括或删除发生时,说明书都被视为含有被修改的组,从而完成了用于所附权利要求的所有马库西群组(Markush group)的书面描述。 When writing any such inclusion or deletion occurs, the specification is considered to be modified containing group, thereby completing all Markush groups (Markush group) for the description of the appended claims.

[0269] 除非另外指出,表达成分的量、性质(例如用于说明书和权利要求的分子量、反应条件等)的所有数字在所有情况下都将被理解为被术语“约”修饰。 All figures [0269] Unless otherwise indicated, expressing quantities of ingredients, properties (e.g., molecular weight used in the specification and claims, reaction conditions, etc.) in each case to be understood as modified by the term "about." 如本文所使用,术语“约” 指如此定量的项目、参数或术语涵盖在规定项目、参数或术语的值上下±10%的范围。 As used herein, the term "about" refers to the amount of such items, parameters, or in terms encompass project a predetermined range, parameter or term value ± 10% of the vertical. 因此,除非指出相反,否则在说明书和所附权利要求中列出的数字参数为近似值,其可根据本发明力图得到的所需性质改变。 Accordingly, unless indicated to the contrary, the numerical parameters set forth in the specification and the appended claims are approximations, the desired properties sought to be obtained which is changed in accordance with the present invention. 至少,并不像试图将等同原则的应用限于权利要求的范围一样,每个数字参数应至少根据报告的有效数位的数字并通过应用普通约数技术来理解。 At least, not as an attempt to apply the doctrine of equivalents as limited to the scope of the claims, each numerical parameter should at least be understood that by applying ordinary rounding techniques according to figures reported significant digits. 尽管列出本发明广泛范围的数值范围和参数为近似值,但在特定实施例中列出的数值都尽可能精确地报告。 Notwithstanding that the numerical ranges and parameters of the broad scope of the invention are approximations, the numerical values ​​set forth in the specific embodiment are reported as precisely as possible. 然而,任何数值固有地含有必定由在其各自的测试测量中发现的标准偏差引起的某些误差。 However, any numerical values ​​inherently contain certain errors necessarily resulting from the standard deviation found in their respective testing measurements.

[0270] 除非本文另外指出或与上下文明显矛盾,用否则于描述本发明的上下文中(尤其是在下面权利要求的上下文中)的术语“一个”、“一种”、“所述”和类似指示物应理解为包括单数和复数。 [0270] Unless otherwise indicated herein or clearly contradicted by context, or used in the context of the present invention (especially in the context of the following claims) describe the term "a", "an", "the" and similar It is to be construed to cover both the singular and the plural. 本文值范围的叙述仅仅旨在用作单独提及属于该范围的每个独立值的速记方法。 Range recited herein are merely intended to serve as a shorthand method of referring individually to each separate value belonging to the range. 除非本文另外指出,否则将每个单独值并入说明书中就如在本文中对其单独叙述一样。 Unless otherwise indicated herein, each individual value is incorporated into the otherwise specification as recited As its individual herein. 除非本文另外指出或另外与上下文明显矛盾,否则本文描述的所有方法可以任何适合顺序进行。 Unless otherwise indicated herein or otherwise clearly contradicted by context otherwise, all methods described herein may be any suitable order. 使用本文提供的任何和所有实施例或示例性的措辞(例如,“例如”)仅旨在更好地说明本发明,而不构成对另外要求保护的本发明范围的限制。 Using any and all embodiments provided herein, the phraseology and examples, or exemplary (e.g., "such as") is intended merely to better illuminate the invention and does not pose a limitation on the scope of the present claims further invention. 不得将说明书中的措辞理解为指出对实施本发明必不可少的任何不要求保护的元素。 Language in the specification shall be understood that any non-claimed element as essential to point out the practice of the invention.

[0271] 可使用由措辞组成或基本上由措辞组成,将本文公开的特定实施方案进一步限制于权利要求中。 [0271] may be used by the language wording or consists essentially of the composition, the particular embodiments disclosed herein further limited in the claims. 当用于权利要求时,无论是作为存档的修正或是每次加入的修正,过渡术语“由......组成”都排除了在权利要求中未指定的任何元素、步骤或成分。 When used in the claims, whether as added each time or archived correction amendment, the transition term "consisting ...... consisting of" excludes any element in all the claims is not specified, step, or ingredient. 过渡术语“基本上由......组成”将权利要求的范围限于特定的材料或步骤和不会实质上影响基本特性和新特性的材料或步骤。 The transition term "consisting essentially of ......" The scope of the claims be limited to the specific materials or steps and materials or steps do not materially affect the basic and novel characteristics of the. 本文固有或明确地描述和启用了如此要求保护的本发明的实施方案。 Herein inherently or expressly described and enabled embodiments of the invention so claimed.

[0272] 在本说明书中引用和鉴定的所有专利、专利公布和其它公布为了描述和公开(例如)在此类公布中描述的可与本发明联合使用的组合物和方法的目的,单独且明确地通过引用整体并入本文。 All patents [0272] and references identified in this specification, patent publications and other publications disclosed and described in order to (e.g.) the purpose of the compositions and methods of the present invention may be used in combination described in such publications, individually and specifically incorporated herein by reference in its entirety. 在本申请的申请日期之前,这些公布仅为它们的公开而提供。 Prior to the filing date of the application, which was only published their open and available. 就这点而言不应理解为承认由于先前的发明或任何其它原因,无权使发明人先于此类公开。 In this regard it should not be construed as an admission by virtue of prior invention or for any other reason, have no right to make such disclosure by the inventor before. 关于日期的所有声明或关于这些文档内容的陈述基于申请人可得的信息,并不构成关于这些文档的日期或内容正确性的任何承认。 All statements as to the date or representation regarding the content of these documents based on the information available to the applicants and does not constitute any admission as to the correctness of the dates or contents of these documents.

Claims (16)

1. 一种可注射的水凝胶组合物用于制备治疗人患者皮肤上的皱纹的药剂的用途,其中所述组合物包含与二胺交联的透明质酸,所述二胺选自赖氨酸、赖氨酸甲酯及其组合,且所述组合物被注射到患者的皮肤。 An injectable hydrogel composition for the medicament of wrinkles on the skin for treating a human patient, wherein said composition comprising hyaluronic acid crosslinked with a diamine, said diamine is selected from Lai histidine, lysine methyl ester, and combinations thereof, and the composition is injected into the skin of the patient.
2. 根据权利要求1所述的用途,其中所述二胺是赖氨酸。 2. The use according to claim 1, wherein said diamine is lysine.
3. 根据权利要求1或2所述的用途,其中所述二胺是赖氨酸甲酯。 3. The use of claim 1 or claim 2, wherein said diamine is lysine methyl ester.
4. 根据权利要求1、2或3所述的用途,其中将所述组合物注射到患者皮肤不大于约1.0mm的深度。 Use according to claim 1, 2 or 3, wherein the composition is injected into the patient's skin is not greater than the depth of about 1.0mm.
5. 根据权利要求4所述的用途,其中将所述组合物注射到患者皮肤不大于约0.8_的深度。 5. The use as claimed in claim 4, wherein the composition is injected into the patient's skin is not greater than the depth of about 0.8_.
6. 根据权利要求5所述的用途,其中将所述组合物注射到患者皮肤不大于约0.6_的深度。 6. Use as claimed in claim 5, wherein the composition is injected into the patient's skin is not greater than the depth of about 0.6_.
7. 根据权利要求1、2或3所述的用途,其中所述组合物具有至多200Pa的储能模量。 7. The use of claim 2 or claim 3, wherein the composition has a storage modulus of at most 200Pa.
8. 根据权利要求7所述的用途,其中所述组合物具有至少25Pa的储能模量。 8. Use according to claim 7, wherein the composition has a storage modulus of at least 25Pa.
9. 根据权利要求7所述的用途,其中所述组合物具有介于约25Pa和约150Pa之间的储能模量。 9. Use according to claim 7, wherein the composition has a storage modulus of between about 25Pa to about 150Pa.
10. 根据权利要求9所述的用途,其中所述组合物具有介于约40Pa和约IOOPa之间的储能模量。 10. Use according to claim 9, wherein the composition has a storage modulus between about IOOPa between about 40Pa.
11. 根据权利要求7所述的用途,其中所述组合物具有不大于约IOOPa的储能模量。 11. Use according to claim 7, wherein the composition has a storage modulus of not greater than about IOOPa.
12. 根据权利要求7所述的用途,其中所述组合物具有不少于约40Pa的储能模量。 12. Use according to claim 7, wherein the composition has a storage modulus of less than about 40Pa of.
13. 根据权利要求1所述的用途,其中所述皱纹是浅表皱纹或细纹。 13. Use according to claim 1, wherein said superficial wrinkles wrinkles or fine lines.
14. 根据权利要求1所述的用途,其中将所述组合物注射到患者皮肤的眶周区、鼻唇沟区、泪沟区或颈部区。 Periorbital region 14. The use according to claim 1, wherein said composition is injected into the skin of the patient, the nasolabial area, the tear trough region or neck region.
15. 根据权利要求1、2或3所述的用途,其中所述水凝胶已被进一步加工以提供约ΙΟμπι 至约ΙΟΟΟμηι的粒度。 15. The use of claim 2 or claim 3, wherein the hydrogel has been further processed to provide from about ΙΟμπι to a particle size of about ΙΟΟΟμηι.
16. 根据权利要求1、2、3或15所述的用途,其中所述组合物还包含未交联的透明质酸聚合物。 16. The use of claim 3 or claim 15, wherein said composition further comprises a non-cross-linked hyaluronic acid polymer.
CN201810249654.6A 2011-08-25 2012-09-13 Dermal filler compositions for fine line treatment CN108379112A (en)

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