WO2020073706A1 - 一种表面修饰的海绵骨针及其制备方法和用途 - Google Patents

一种表面修饰的海绵骨针及其制备方法和用途 Download PDF

Info

Publication number
WO2020073706A1
WO2020073706A1 PCT/CN2019/096166 CN2019096166W WO2020073706A1 WO 2020073706 A1 WO2020073706 A1 WO 2020073706A1 CN 2019096166 W CN2019096166 W CN 2019096166W WO 2020073706 A1 WO2020073706 A1 WO 2020073706A1
Authority
WO
WIPO (PCT)
Prior art keywords
group
bone needle
sponge bone
modified
hydrocarbon group
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/CN2019/096166
Other languages
English (en)
French (fr)
Inventor
陈铭
张柯达
王德祥
赵婧华
张驰
欧徽龙
张赛曼
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Shenzhen C&d Bio Pharm Co Ltd
Xiamen University
Original Assignee
Shenzhen C&d Bio Pharm Co Ltd
Xiamen University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Shenzhen C&d Bio Pharm Co Ltd, Xiamen University filed Critical Shenzhen C&d Bio Pharm Co Ltd
Publication of WO2020073706A1 publication Critical patent/WO2020073706A1/zh
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/24—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing atoms other than carbon, hydrogen, oxygen, halogen, nitrogen or sulfur, e.g. cyclomethicone or phospholipids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/58—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing atoms other than carbon, hydrogen, halogen, oxygen, nitrogen, sulfur or phosphorus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/58—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing atoms other than carbon, hydrogen, halogen, oxygen, nitrogen, sulfur or phosphorus
    • A61K8/585—Organosilicon compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M37/00—Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • C—CHEMISTRY; METALLURGY
    • C01—INORGANIC CHEMISTRY
    • C01B—NON-METALLIC ELEMENTS; COMPOUNDS THEREOF; METALLOIDS OR COMPOUNDS THEREOF NOT COVERED BY SUBCLASS C01C
    • C01B33/00—Silicon; Compounds thereof
    • C01B33/113—Silicon oxides; Hydrates thereof
    • C01B33/12—Silica; Hydrates thereof, e.g. lepidoic silicic acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/10—General cosmetic use

Definitions

  • the invention belongs to a sponge bone needle penetration-promoting technology, and in particular relates to a surface modified sponge bone needle and a preparation method and application thereof.
  • Sponge bone needles are the siliceous or calcareous fibrous materials that support the skeleton and resist external invasion in the sponge. They have various structural forms, including uniaxial, biaxial, triaxial and multiaxial. The study found that the sponge bone needle has good optical properties and mechanical properties. At present, the research on the sponge bone needle focuses on the optical properties, nanostructure, bionics, and medical transdermal agents of the sponge bone needle. Some natural spongy bone needles with high mechanical strength and sharp ends have a role similar to microneedles, which can stay in the stratum corneum of the skin for a long time and form a large number of persistent microchannels.
  • the spongy bone needles are used alone or with some
  • the combined application of carriers can effectively promote the transdermal absorption of drugs or active ingredients, especially suitable for active ingredients of drugs or cosmetics with small molecular weight and good stability, but for some large molecules, it is necessary to develop some more effective ways to promote penetration .
  • the purpose of the present invention is to overcome the shortcomings of the prior art, and to provide a surface modified sponge bone needle and its preparation method and use.
  • a surface-modified sponge bone needle the surface of the sponge bone needle is modified with an aliphatic hydrocarbon group, an aromatic hydrocarbon group, an aramid group or other organic groups derived based on these three types of groups.
  • the aliphatic hydrocarbon group, aromatic hydrocarbon group, aryl fat group or derivative group thereof is combined with the sponge bone needle through a -Si-O-Si- bond, and the aliphatic hydrocarbon group, aromatic hydrocarbon group, aryl fat group or derivative thereof
  • the groups can be connected to each other through a -Si-O-Si- bond, thereby forming a thin film of aliphatic hydrocarbon group, aromatic hydrocarbon group, aryl fatty group or derivative group on the surface of the sponge bone needle.
  • the structural formula of the surface-modified sponge bone needle is shown in the following formula I:
  • R is the aforementioned aliphatic hydrocarbon group, aromatic hydrocarbon group, arylaliphatic group or a derivative group thereof.
  • the aromatic hydrocarbon group represents that the aromatic ring in R is directly connected to Si, and there may be other substituents on the aromatic ring.
  • the arylaliphatic group represents that the aromatic ring in R is not directly connected to Si, and there are groups such as aliphatic hydrocarbon groups between the aromatic ring and Si, and there may be other substituents on the aromatic ring.
  • the sponge bone needle is derived from the bee sponge Haliclona sp ..
  • the aliphatic hydrocarbon group is a C3-C18 alkyl group.
  • the aromatic hydrocarbon group is phenyl
  • the arylaliphatic group is benzyl or phenethyl.
  • a preparation method of surface modified sponge bone needles includes:
  • step 2) Add the surface silanol group-activated sponge bone needle obtained in step 1) to an anhydrous aprotic solvent, add a silane coupling agent at 80 to 100 ° C, the sponge bone needle and the silane coupling agent
  • the mass ratio is 1: 0.5 to 2
  • the concentration of the silane coupling agent in the anhydrous aprotic solvent is 1 to 50 mM; after the reaction for 2 to 12 hours, a surface-modified sponge bone needle is prepared.
  • the method further includes: ultrasonically cleaning and drying the sponge bone needle with water, ethanol, and acetone in sequence to obtain a sponge bone needle with a clean surface; and then adding the NaOH solution and adding the sponge bone needle with the clean surface Silanol groups on the surface of hydrochloric acid solution, ultrasound, filtration, washing and drying are activated.
  • the purity of the sponge bone needle is not less than 90%.
  • the sponge bone needle has a uniform shape and a complete structure.
  • the anhydrous aprotic solvent is anhydrous toluene.
  • the silane coupling agent is of the general formula R-SiX 3 , R-Si (R ′) 2 -X or R-Si (R ′)-X 2 ;
  • X is an alkoxy group (eg Methoxy, ethoxy, isopropoxy, etc.), acyloxy (such as acetoxy, etc.) or halo (such as chlorine substitution, etc.);
  • the R is the above aliphatic hydrocarbon group (such as C3-C18 Alkyl, etc.), aromatic hydrocarbon groups (such as phenyl, etc.), araliphatic groups (such as benzyl, phenethyl, etc.) or derivatives thereof;
  • the R ′ is a short-chain aliphatic hydrocarbon group (short-chain aliphatic hydrocarbon group generally refers to It is a C1-C3 hydrocarbon group, such as methyl, ethyl, etc.).
  • the sponge bone needle itself has a silanol group (-SiOH), after the silanol group is activated, -Si-OH reacts with the R-Si-X of the silane coupling agent to form -Si (formula (Not shown in I) -O-Si-R, R (that is, aliphatic hydrocarbon group, aromatic hydrocarbon group, aryl aliphatic group or derivative group thereof) is bonded to the sponge bone needle through a -Si-O-Si- bond.
  • R-Si- can also form -Si-O-Si- bonds to connect with each other.
  • the specific silane coupling agent is shown in the following table:
  • the surface modified sponge bone needle can absorb drugs, fluorescent molecules, polymers, proteins, peptides, nanoparticles through physical action
  • the microneedle-like action of the sponge bone needle itself can be used to promote the percutaneous absorption or skin absorption of the above substances, and can be used to promote the absorption of drugs, cosmetics and the like.
  • the transdermal absorption composition refers to the medicine absorbed from the skin into the systemic blood circulation and reaching the effective blood drug concentration to achieve disease treatment or prevention, that is, systemic administration and systemic administration; the skin absorption
  • the composition refers to the way in which the drug reaches an effective concentration locally on the skin to prevent or treat disease, that is, a local administration method.
  • the synthesis process of the invention is simple, the reaction conditions are easy to control, and the physical and chemical properties of the product are stable.
  • the product can physically adsorb one or more polypeptides or proteins, and then promote the skin or transdermal administration of the polypeptides and proteins through microneedle-like action.
  • the penetration of macromolecular substances into the skin provides an effective way, with broad application prospects and huge potential market value.
  • Fig. 1 is a fluorescence micrograph of GFP (green fluorescent protein) adsorbed by a bee sponge bone needle after silane modification.
  • A Fluorescence image of unmodified bone needle in the control group
  • B fluorescence image of butyl trimethoxysilane modified bee sponge bone needle adsorbing GFP
  • C fluorescence image of octyl trimethoxysilane modified bee sponge bone needle adsorbing GFP.
  • the scale in the figure is 100 ⁇ m.
  • Figure 2 is a quantitative comparison of transdermal results of GFP direct transdermal, unmodified bone needles after transdermal and silane-modified bee sponge bone needles adsorbing the same amount of GFP after transdermal massage.
  • Fig. 3 is a comparison chart of fluorescent slices after GFP direct transdermal, unmodified bone needle massage after transdermal and silane-modified bee sponge bone needles adsorbing the same amount of GFP massage.
  • D modification of octyltrimethoxysilane modification group Fluorescence image of tissue section.
  • the scale in the figure is 100 ⁇ m.
  • step (2) Add the cleaned bee sponge bone needle obtained in step (1) to a 5 mol / L NaOH solution and sonicate for 30 min; after filtering, washing and drying, add the sponge bone needle to a 10% hydrochloric acid solution and sonicate After 30 minutes, filter, wash and dry to obtain the bee sponge bone needle activated by silanol groups on the surface;
  • step (3) Add the surface silanol group-activated bee sponge bone needle obtained in step (2) to anhydrous toluene, add butyltrimethoxysilane at 90 ° C, the sponge bone needle and butyltrimethoxy group
  • the mass ratio of silane is 1: 1, and the concentration of butyltrimethoxysilane in anhydrous toluene is 10mM; after 8 hours of reaction, a butyltrimethoxysilane modified bee sponge bone needle is prepared.
  • step (2) Add the cleaned bee sponge bone needle obtained in step (1) to a 10 mol / L NaOH solution and sonicate for 10 min; after filtering, washing and drying, add the sponge bone needle to a 20% hydrochloric acid solution and sonicate 10min, filter, wash and dry to obtain bee sponge bone needle activated by silanol group on the surface;
  • step (3) Add the surface silanol group-activated bee sponge bone needle obtained in step (2) to anhydrous toluene, add octyltrimethoxysilane at 95 ° C, the sponge bone needle and octyltrimethoxy group
  • the mass ratio of silane is 1: 2, and the concentration of octyltrimethoxysilane in anhydrous toluene is 5mM; after reaction for 12h, octyltrimethoxysilane modified bee sponge bone needles are prepared.
  • Example 3 Confirmation of butyl trimethoxysilane modified bee sponge bone needle and octyl trimethoxysilane modified bee sponge bone needle
  • XPS X-ray photoelectron spectroscopy
  • Example 4 Application of butyltrimethoxysilane modified bee sponge bone needle and octyltrimethoxysilane modified bee sponge bone needle
  • the mass of GFP adsorbed by butyl trimethoxysilane modified bee sponge bone needle and octyl trimethoxysilane modified bee sponge bone needle were 1.1 ⁇ g / mg and 1.4 ⁇ g / mg, respectively.
  • the bone needles of each group in (1) mixed with GFP solution, filtered and washed were directly smeared on the isolated pig skin, and the GFP solution was directly smeared on the isolated pig skin as a control.
  • Each group cooperated with massage to assist absorption. After 8 hours, the distribution of green fluorescent protein in each layer of the skin is shown in Figure 3, and the quantitative comparison is shown in Figure 2.
  • the GFP fluorescence value in the skin was about 700 (bone needle group in Figure 2), It indicates that the bee sponge bone needle without surface modification can also promote the transdermal absorption of GFP to a certain extent, which may be caused by the microneedle effect of the bee sponge bone needle itself, but the GFP molecular weight is large, and the tissue section is placed Observed under a fluorescence microscope, it can be seen that GFP is basically located in the superficial layer of the stratum corneum of the skin, which is difficult to absorb deeply (Figure 3B).
  • the GFP fluorescence value in the skin can reach more than 1200 (Figure 2 butyl modified group And octyl-modified group), tissue sections were observed under a fluorescent microscope, and the epidermal layer and dermal layer of the skin had green fluorescence ( Figure 3B and Figure 3D).
  • the surface modification of the bee sponge bone needle it can adsorb GFP and It penetrates into the skin and releases it to promote the transdermal absorption and depth of GFP.
  • the invention provides a surface-modified sponge bone needle and its preparation method and use.
  • the synthesis process of the invention is simple, the reaction conditions are easy to control, and the physical and chemical properties of the product are stable.
  • the product can physically adsorb one or more polypeptides or proteins.
  • the microneedle-like action promotes the skin or transdermal administration of peptides and proteins, and provides an effective way for the penetration of macromolecular substances into the skin. It has broad application prospects, huge potential market value, and good industrial practicality.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Epidemiology (AREA)
  • Dermatology (AREA)
  • Medicinal Chemistry (AREA)
  • General Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Birds (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Biophysics (AREA)
  • Molecular Biology (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Hematology (AREA)
  • Anesthesiology (AREA)
  • Medical Informatics (AREA)
  • Biomedical Technology (AREA)
  • Inorganic Chemistry (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Medicinal Preparation (AREA)
  • Cosmetics (AREA)
  • Materials For Medical Uses (AREA)

Abstract

一种表面修饰脂肪烃基、芳烃基、芳脂基或其衍生基团的海绵骨针，所述海绵骨针的制备方法为：对海绵骨针进行清洗去除杂质；活化海绵骨针表面的硅醇基；再将其与硅烷偶联剂反应而得。所述海绵骨针可物理吸附一种或多种多肽或蛋白质，进而通过微针作用促进多肽和蛋白质的皮肤或经皮给药。

Description

一种表面修饰的海绵骨针及其制备方法和用途 技术领域
本发明属于海绵骨针促渗技术，具体涉及一种表面修饰的海绵骨针及其制备方法和用途。
背景技术
海绵骨针(Sponge spicules)是海绵中起骨架支撑和抵御外侵的硅质或钙质纤维状物，具有多种结构形态，包括单轴、双轴、三轴和多轴等。研究发现，海绵骨针具有良好的光学特性和力学性能，目前关于海绵骨针的研究集中在海绵骨针的光学特性、纳米结构、仿生学和医药透皮剂等领域。一些天然具有较高的机械强度、末端尖锐的海绵骨针，具有类似于微针的作用，可长时间滞留于皮肤角质层并形成大量持续存在的微通道，海绵骨针单独应用，或与一些载体联合应用，可以有效促进药物或活性成分的经皮吸收，特别适合分子量较小且稳定性较好的药物或化妆品活性成分，但针对一些大分子，还需要开发出一些更有效的促渗途径。
发明内容
本发明的目的在于克服现有技术的不足之处，提供了一种表面修饰的海绵骨针及其制备方法和用途。
本发明解决其技术问题所采用的技术方案之一是：
一种表面修饰的海绵骨针，所述海绵骨针表面修饰有脂肪烃基、芳烃基、芳脂基或基于这三类基团衍生出的其他有机基团。
一实施例中：所述脂肪烃基、芳烃基、芳脂基或其衍生基团与海绵骨针通过-Si-O-Si-键结合，所述脂肪烃基、芳烃基、芳脂基或其衍生基团可通过-Si-O-Si-键相互连接，从而在海绵骨针表面形成一层脂肪烃基、芳烃基、芳脂基或其衍生基团薄膜。
一实施例中：所述表面修饰的海绵骨针的结构式如下式Ⅰ所示：
Figure PCTCN2019096166-appb-000001
式Ⅰ
其中，R为所述脂肪烃基、芳烃基、芳脂基或其衍生基团。
其中，所述芳烃基代表R中的芳香环直接与Si相连，且该芳香环上还可以有其他取代基。所述芳脂基代表R中的芳香环未直接与Si相连，芳香环与Si之间还有脂肪烃基等基团，且该芳香环上也还可以有其他取代基。
一实施例中：所述海绵骨针来源于蜂海绵Haliclona sp.。
一实施例中：所述脂肪烃基为C3～C18的烷基。
一实施例中：所述芳烃基为苯基。
一实施例中：所述芳脂基为苯甲基或苯乙基。
本发明解决其技术问题所采用的技术方案之二是：
一种表面修饰的海绵骨针的制备方法，包括：
1)将海绵骨针加入1～10mol/L的NaOH溶液中，超声10～60min；过滤、洗涤、干燥后，再将海绵骨针加入到5～20％盐酸溶液中，超声10～60min，过滤、洗涤、干燥，得到表面硅醇基活化的海绵骨针；
2)将步骤1)得到的表面硅醇基活化的海绵骨针加入至无水非质子性溶剂中，80～100℃下，加入硅烷偶联剂，所述海绵骨针与硅烷偶联剂的质量比为1：0.5～2，所述硅烷偶联剂在无水非质子性溶剂中的浓度为1～50mM；反应2～12h后，制得表面修饰的海绵骨针。
一实施例中：还包括：依次采用水、乙醇、丙酮对海绵骨针进行超声清洗、干燥，得到表面清洁的海绵骨针；然后将该表面清洁的海绵骨针进行所述加入NaOH溶液、加入盐酸溶液、超声、过滤、洗涤、干燥的表面硅醇基活化。
一实施例中：所述海绵骨针的纯度不低于90％，最好，海绵骨针形态均一、结构 完整。
一实施例中：所述无水非质子性溶剂为无水甲苯。
一实施例中：所述硅烷偶联剂为通式R-SiX 3、R-Si(R’) 2-X或R-Si(R’)-X 2；所述X为烷氧基(例如甲氧基、乙氧基、异丙氧基等)、酰氧基(例如乙酰氧基等)或卤代基(例如氯取代等)；所述R为上述的脂肪烃基(例如C3～C18的烷基等)、芳烃基(例如苯基等)、芳脂基(例如苯甲基、苯乙基等)或其衍生基团；所述R’为短链脂肪烃基(短链脂肪烃基一般指的是C1～C3的烃基，例如甲基、乙基等)等。
所述制备方法的原理为：海绵骨针本身具有硅醇基(-SiOH)，硅醇基经活化后，-Si-O-H与硅烷偶联剂的R-Si-X反应，形成-Si(式Ⅰ中未示出)-O-Si-R，R(即脂肪烃基、芳烃基、芳脂基或其衍生基团)通过-Si-O-Si-键与海绵骨针结合。R-Si-之间也可形成-Si-O-Si-键相互连接。
一实施例中：具体的硅烷偶联剂如下表所示：
Figure PCTCN2019096166-appb-000002
本发明解决其技术问题所采用的技术方案之三是：
一种表面修饰的海绵骨针在制备经皮吸收组合物或皮肤吸收组合物中的用途，该表面修饰的海绵骨针可通过物理作用吸附药物、荧光分子、聚合物、蛋白质、多肽、纳米颗粒等，进而利用海绵骨针本身的类微针作用促进上述物质的经皮吸收或皮肤吸收，可用于促进药物、化妆品等的吸收。
其中，所述经皮吸收组合物指的是用药后，药物由皮肤吸收进入全身血液循环并达到有效血药浓度、实现疾病治疗或预防，即系统给药、全身给药方式；所述皮肤吸收组合物指的是用药后，药物在皮肤局部达到有效浓度，实现疾病预防或治疗，即局部给药方式。
本技术方案与背景技术相比，它具有如下优点：
本发明的合成工艺简单，反应条件易控制，产品物化性质稳定，该产品可物理吸附一种或多种多肽或蛋白质，进而通过类微针作用促进多肽和蛋白质的皮肤或经皮给药，为大分子物质渗透进皮肤提供了有效途径，应用前景广泛，潜在市场价值巨大。
附图说明
下面结合附图和实施例对本发明作进一步说明。
图1为硅烷基修饰后的蜂海绵骨针吸附GFP(绿色荧光蛋白)的荧光显微镜图。A：对照组未修饰骨针的荧光图，B：丁基三甲氧基硅烷修饰蜂海绵骨针吸附GFP的荧光图，C：辛基三甲氧基硅烷修饰蜂海绵骨针吸附GFP的荧光图。图中标尺为100μm。
图2为GFP直接透皮、未修饰骨针按摩后透皮与硅烷基修饰蜂海绵骨针吸附等量GFP按摩后透皮结果定量对比图。
图3为GFP直接透皮、未修饰骨针按摩后透皮与硅烷基修饰蜂海绵骨针吸附等量GFP按摩后的荧光切片对比图。A：GFP直接透皮对照组组织切片荧光图，B：未修饰 骨针组组织切片荧光图，C：丁基三甲氧基硅烷修饰组组织切片荧光图，D：辛基三甲氧基硅烷修饰组组织切片荧光图。图中标尺为100μm。
具体实施方式
下面通过实施例具体说明本发明的内容：
实施例1：丁基三甲氧基硅烷修饰蜂海绵骨针的制备
(1)依次采用水、乙醇、丙酮对蜂海绵骨针进行超声清洗、干燥，得到表面清洁的蜂海绵骨针。
(2)将步骤(1)得到的表面清洁的蜂海绵骨针加入5mol/L的NaOH溶液中，超声30min；过滤、洗涤、干燥后，再将海绵骨针加入到10％盐酸溶液中，超声30min，过滤、洗涤、干燥，得到表面硅醇基活化的蜂海绵骨针；
(3)将步骤(2)得到的表面硅醇基活化的蜂海绵骨针加入至无水甲苯中，90℃下，加入丁基三甲氧基硅烷，所述海绵骨针与丁基三甲氧基硅烷的质量比为1：1，丁基三甲氧基硅烷在无水甲苯中的浓度为10mM；反应8h后，制得丁基三甲氧基硅烷修饰蜂海绵骨针。
实施例2：辛基三甲氧基硅烷修饰蜂海绵骨针的制备
(1)依次采用水、乙醇、丙酮对蜂海绵骨针进行超声清洗、干燥，得到表面清洁的蜂海绵骨针。
(2)将步骤(1)得到的表面清洁的蜂海绵骨针加入10mol/L的NaOH溶液中，超声10min；过滤、洗涤、干燥后，再将海绵骨针加入到20％盐酸溶液中，超声10min，过滤、洗涤、干燥，得到表面硅醇基活化的蜂海绵骨针；
(3)将步骤(2)得到的表面硅醇基活化的蜂海绵骨针加入至无水甲苯中，95℃下，加入辛基三甲氧基硅烷，所述海绵骨针与辛基三甲氧基硅烷的质量比为1：2，辛 基三甲氧基硅烷在无水甲苯中的浓度为5mM；反应12h后，制得辛基三甲氧基硅烷修饰蜂海绵骨针。
实施例3：丁基三甲氧基硅烷修饰蜂海绵骨针、辛基三甲氧基硅烷修饰蜂海绵骨针的确证
丁基三甲氧基硅烷和辛基三甲氧基硅烷表面修饰的蜂海绵骨针的X射线光电子能谱(XPS)元素分析结果如下：
Figure PCTCN2019096166-appb-000003
从上表中可知，采用丁基三甲氧基硅烷、辛基三甲氧基硅烷分别对蜂海绵骨针进行表面修饰后，与未修饰的骨针相比，C元素比例升高，说明丁基和辛基已连接到蜂海绵骨针表面，即丁基三甲氧基硅烷修饰的蜂海绵骨针及辛基三甲氧基硅烷修饰的蜂海绵骨针的制备成功。
实施例4：丁基三甲氧基硅烷修饰蜂海绵骨针、辛基三甲氧基硅烷修饰蜂海绵骨针的应用
(1)未修饰与表面修饰的蜂海绵骨针对GFP的吸附作用
将实施例1制得的丁基三甲氧基硅烷修饰蜂海绵骨针与绿色荧光蛋白溶液混合，通过物理作用进行蛋白吸附；将实施例2制得的辛基三甲氧基硅烷修饰蜂海绵骨针与绿色荧光蛋白溶液混合，通过物理作用进行蛋白吸附；同时，采用未进行表面修饰的蜂海绵骨针与同样的绿色荧光蛋白溶液混合。混合1小时后，过滤、洗涤，将各组骨针置于荧光显微镜下观察，结果如附图1所示。
从图1中可知，未进行表面修饰的蜂海绵骨针，过滤洗涤后将其置于荧光显微镜下观察时(如图1A)，几乎没有荧光显示，表明未进行表面修饰的蜂海绵骨针几乎无法吸附GFP或吸附量很低；而丁基三甲氧基硅烷修饰蜂海绵骨针与辛基三甲氧基硅烷修饰蜂海绵骨针均可吸附GFP，过滤洗涤后将其置于荧光显微镜下观察可见呈现出明显的绿色荧光(如图1B与图1C)，表明丁基三甲氧基硅烷修饰蜂海绵骨针与辛基三甲氧基硅烷修饰蜂海绵骨针均已吸附大量的GFP。经进一步计算，丁基三甲氧基硅烷修饰蜂海绵骨针与辛基三甲氧基硅烷修饰蜂海绵骨针吸附GFP的质量分别为1.1μg/mg和1.4μg/mg。
(2)未修饰与表面修饰的蜂海绵骨针对GFP的促进透皮作用
将(1)中与GFP溶液混合、过滤、洗涤后的各组骨针直接涂抹于离体猪皮上，同时将GFP溶液直接涂抹在离体猪皮上作为对照，各组配合按摩辅助吸收。8小时后绿色荧光蛋白在皮肤各层的分布情况见附图3，定量对比如附图2。
结合图2与图3可知，GFP溶液直接涂抹并辅助按摩吸收后，皮肤内GFP荧光值小于400(图2中对照组)，组织切片置于荧光显微镜下观察皮肤内几乎无绿色荧光(图3A)，表明直接涂抹按摩GFP几乎无法实现透皮吸收。未进行表面修饰的蜂海绵骨针与同样的绿色荧光蛋白溶液混合一段时间后，过滤，洗涤，直接涂抹并辅助按摩吸收后，皮肤内GFP荧光值约为700(图2中骨针组)，说明未进行表面修饰的蜂海绵骨针也可在一定程度上促进GFP透皮吸收，可能是通过蜂海绵骨针本身的类微针效应带来的效果，但GFP分子量较大，组织切片置于荧光显微镜下观察，可见GFP基本位于皮肤的角质层浅表层，难以深层吸收(图3B)。而丁基三甲氧基硅烷修饰蜂海绵骨针与辛基三甲氧基硅烷修饰蜂海绵骨针直接涂抹并辅助按摩吸收后，皮肤内GFP荧光值均可达1200以上(图2中丁基修饰组与辛基修饰组)，组织切片置于荧光显微镜下观察， 在皮肤的表皮层、真皮层均有绿色荧光(图3B与图3D)，表面蜂海绵骨针进行表面修饰后，可吸附GFP并刺入皮下释放，促进GFP的透皮吸收量与透皮深度。
以上所述，仅为本发明较佳实施例而已，故不能依此限定本发明实施的范围，即依本发明专利范围及说明书内容所作的等效变化与修饰，皆应仍属本发明涵盖的范围内。
工业实用性
本发明提供了一种表面修饰的海绵骨针及其制备方法和用途，本发明的合成工艺简单，反应条件易控制，产品物化性质稳定，该产品可物理吸附一种或多种多肽或蛋白质，进而通过类微针作用促进多肽和蛋白质的皮肤或经皮给药，为大分子物质渗透进皮肤提供了有效途径，应用前景广泛，潜在市场价值巨大,具有良好的工业实用性。

Claims (13)

  1. 一种表面修饰的海绵骨针，其特征在于：所述海绵骨针表面修饰有脂肪烃基、芳烃基、芳脂基或基于这三类基团衍生出的其他有机基团。
  2. 根据权利要求1所述的表面修饰的海绵骨针，其特征在于：所述脂肪烃基、芳烃基、芳脂基或其衍生基团与海绵骨针通过-Si-O-Si-键结合；所述脂肪烃基、芳烃基、芳脂基或其衍生基团通过-Si-O-Si-键相互连接。
  3. 根据权利要求1所述的表面修饰的海绵骨针，其特征在于：所述表面修饰的海绵骨针的结构式如下式所示：
    Figure PCTCN2019096166-appb-100001
    其中，R为所述脂肪烃基、芳烃基、芳脂基或其衍生基团。
  4. 根据权利要求1所述的表面修饰的海绵骨针，其特征在于：所述海绵骨针来源于蜂海绵Haliclona sp.；所述脂肪烃基为C3～C18的烷基；所述芳烃基为苯基；所述芳脂基为苯甲基或苯乙基。
  5. 根据权利要求1所述的表面修饰的海绵骨针，其特征在于：所述海绵骨针表面修饰硅烷偶联剂的基团。
  6. 一种权利要求1至5中任一项所述的表面修饰的海绵骨针的制备方法，其特征在于：包括：
    1)将海绵骨针加入1～10mol/L的NaOH溶液中，超声10～60min；过滤、洗涤、干燥后，再将海绵骨针加入到5～20％盐酸溶液中，超声10～60min，过滤、洗涤、干燥，得到表面硅醇基活化的海绵骨针；
    2)将步骤1)得到的表面硅醇基活化的海绵骨针加入至无水非质子性溶剂中，80～100℃下，加入硅烷偶联剂，所述海绵骨针与硅烷偶联剂的质量比为1：0.5～2，所述硅烷偶联剂在无水非质子性溶剂中的浓度为1～50mM；反应2～12h后，制得表面修饰的海绵骨针。
  7. 根据权利要求6所述的制备方法，其特征在于：还包括：依次采用水、乙醇、丙酮对海绵骨针进行超声清洗、干燥，得到表面清洁的海绵骨针；然后将该表面清洁的海绵骨针进行所述表面硅醇基活化。
  8. 根据权利要求6所述的制备方法，其特征在于：所述无水非质子性溶剂为无水甲苯。
  9. 根据权利要求6所述的制备方法，其特征在于：所述硅烷偶联剂为通式R-SiX 3、R-Si(R’) 2-X或R-Si(R’)-X 2；所述X为烷氧基、酰氧基或卤代基；所述R为所述脂肪烃基、芳烃基、芳脂基或其衍生基团；所述R’为短链脂肪烃基。
  10. 根据权利要求9所述的制备方法，其特征在于：所述烷氧基为甲氧基、乙氧基或异丙氧基；所述酰氧基为乙酰氧基；所述卤代基为氯取代；所述短链脂肪烃基为甲基或乙基。
  11. 根据权利要求6所述的制备方法，其特征在于：所述硅烷偶联剂为丁基三甲氧基硅烷，丁基三乙氧基硅烷，辛基三甲氧基硅烷，苯甲基三氯硅烷，苯乙基三乙氧基硅烷，苯基三甲氧基硅烷，十二烷基甲基二乙氧基硅烷，辛基甲基二甲氧基硅烷，癸基甲基二氯硅烷，己基二甲基乙酰氧基硅烷，十八烷基二甲基氯代硅烷，辛基二甲基甲氧基硅烷。
  12. 根据权利要求6所述的制备方法，其特征在于：所述海绵骨针的纯度不低于90％。
  13. 一种权利要求1至5中任一项所述的表面修饰的海绵骨针在制备经皮吸收组合物或皮肤吸收组合物中的用途。
PCT/CN2019/096166 2018-10-09 2019-07-16 一种表面修饰的海绵骨针及其制备方法和用途 Ceased WO2020073706A1 (zh)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201811175149.8 2018-10-09
CN201811175149.8A CN111012916B (zh) 2018-10-09 2018-10-09 一种表面修饰的海绵骨针及其制备方法和用途

Publications (1)

Publication Number Publication Date
WO2020073706A1 true WO2020073706A1 (zh) 2020-04-16

Family

ID=70165072

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2019/096166 Ceased WO2020073706A1 (zh) 2018-10-09 2019-07-16 一种表面修饰的海绵骨针及其制备方法和用途

Country Status (2)

Country Link
CN (1) CN111012916B (zh)
WO (1) WO2020073706A1 (zh)

Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109381422B (zh) * 2017-08-07 2020-11-06 厦门大学 一种经皮吸收组合物及其在制备经皮吸收制剂中的用途
CN112603880A (zh) * 2020-12-21 2021-04-06 梁丽君 表皮重建制剂的制备方法与应用以及表皮重建制剂与使用方法
CN115400068B (zh) * 2022-09-30 2024-05-17 广州吉腾生物科技有限公司 一种修护舒缓组合物及含有该组合物的水及乳及霜
CN117100682A (zh) * 2023-05-12 2023-11-24 厦门大学 中空海绵骨针及其制备方法和应用
CN119386280B (zh) * 2024-11-05 2026-01-30 西南交通大学 一种高渗透药物涂层球囊及其制备方法和应用
CN119524155B (zh) * 2024-11-29 2026-03-27 厦门大学 一种表面吸附纳米颗粒的海绵骨针及其制备方法和用途

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101130764A (zh) * 2006-08-23 2008-02-27 中国科学院大连化学物理研究所 反应器培养海绵或其细胞生产生物硅质材料的方法及装置
CN105858669A (zh) * 2016-04-27 2016-08-17 厦门大学 高纯度海绵骨针的制备方法
CN105999535A (zh) * 2016-04-27 2016-10-12 厦门大学 海绵骨针作为皮肤物理促渗剂的应用
CN108163861A (zh) * 2018-02-09 2018-06-15 四川理工学院 一种硅质海绵骨针的纯化方法和纯化系统及应用

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1289184C (zh) * 2004-06-02 2006-12-13 武汉大学 环糊精键合硅胶固定相及其制备方法和用途
JP6163486B2 (ja) * 2012-07-02 2017-07-12 大日本住友製薬株式会社 癌抗原ペプチド経皮剤
CN114515269A (zh) * 2015-04-06 2022-05-20 株式会社Lg生活健康 微针及其制造方法

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101130764A (zh) * 2006-08-23 2008-02-27 中国科学院大连化学物理研究所 反应器培养海绵或其细胞生产生物硅质材料的方法及装置
CN105858669A (zh) * 2016-04-27 2016-08-17 厦门大学 高纯度海绵骨针的制备方法
CN105999535A (zh) * 2016-04-27 2016-10-12 厦门大学 海绵骨针作为皮肤物理促渗剂的应用
CN108163861A (zh) * 2018-02-09 2018-06-15 四川理工学院 一种硅质海绵骨针的纯化方法和纯化系统及应用

Also Published As

Publication number Publication date
CN111012916A (zh) 2020-04-17
CN111012916B (zh) 2021-10-01

Similar Documents

Publication Publication Date Title
WO2020073706A1 (zh) 一种表面修饰的海绵骨针及其制备方法和用途
Hu et al. Mechanically active adhesive and immune regulative dressings for wound closure
Deng et al. Lamprey-teeth-inspired oriented antibacterial sericin microneedles for infected wound healing improvement
Nyström et al. Ice-templated and cross-linked amyloid fibril aerogel scaffolds for cell growth
Jin et al. Multifunctional self-healing peptide hydrogel for wound healing
US20190202998A1 (en) Visible light-curable water-soluble chitosan derivative, chitosan hydrogel, and preparation method therefor
JP5932824B2 (ja) グルカン
US10836872B2 (en) Visible light-curable water-soluble chitosan derivative, chitosan hydrogel, and preparation method therefor
BR112012026330B1 (pt) composição antimicrobiana, bem como gel e curativo antimicrobianos, seus usos e métodos de preparação
CN105451745A (zh) 含有阳离子化壳聚糖的药物
CN110812688A (zh) 一种透皮给药的微针及其制备方法
Pinese et al. Bioactive peptides grafted silicone dressings: A simple and specific method
CN109152794A (zh) 包含在哺乳动物组织中增加氧水平的包合物的聚氧化金属氢氧化物
Chen et al. Multifunctional layered microneedle patches enable transdermal angiogenesis and immunomodulation for scarless healing of thermal burn injuries
Chen et al. Effective scald wound functional recovery patch achieved by molecularly intertwined electrical and chemical message in self-adhesive assemblies
Lim et al. Bioinspired suction-driven strategies with nanoscale skin-controllable adhesive architectures for efficient liquid formulated transdermal patches
Zhang et al. A bilayer polyurethane patch with sustained growth factor release and antibacteria for re-epithelization of large-scale oral mucosal defects
WO2024235113A1 (zh) 中空海绵骨针及其制备方法和应用
CN108653745B (zh) 一种透明质酸前药及其制备方法和在透皮给药中的应用
Hou et al. Preparation of Sodium Alginate Hydrogels with Bioadhesive and Photothermal Effects to Suppress Bacterial Infection and Promote Wound Healing
CN113975249A (zh) Tris-BNP纳米颗粒的制备及其在皮肤疾病治疗中的应用
CN114834066B (zh) 复合多层微针的制备方法
CN107190300A (zh) 介孔羟基磷灰石/二氧化钛纳米管阵列复合材料的制备方法
CN121221568B (zh) 一种负载山楂酸的复合纳米压电材料外敷贴及其制备方法与应用
Zeng et al. Hydrophobic sponge spicule-mediated transdermal vaccination combines controlled skin inflammation as self-adjuvant with dual immune activation

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 19871488

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 19871488

Country of ref document: EP

Kind code of ref document: A1