WO2018226160A1 - Patch à micro-aiguilles chargé d'un agent de brunissement de graisse et son procédé de préparation - Google Patents

Patch à micro-aiguilles chargé d'un agent de brunissement de graisse et son procédé de préparation Download PDF

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Publication number
WO2018226160A1
WO2018226160A1 PCT/SG2018/050282 SG2018050282W WO2018226160A1 WO 2018226160 A1 WO2018226160 A1 WO 2018226160A1 SG 2018050282 W SG2018050282 W SG 2018050282W WO 2018226160 A1 WO2018226160 A1 WO 2018226160A1
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microneedle
microneedles
lactic
poly
microneedle patch
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PCT/SG2018/050282
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English (en)
Inventor
Peng Chen
Chenjie Xu
Aung THAN
Fengna XI
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Pointed Biotech Pte Ltd
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Publication of WO2018226160A1 publication Critical patent/WO2018226160A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/357—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having two or more oxygen atoms in the same ring, e.g. crown ethers, guanadrel
    • A61K31/36—Compounds containing methylenedioxyphenyl groups, e.g. sesamin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • A61K9/0021—Intradermal administration, e.g. through microneedle arrays, needleless injectors
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/195—Carboxylic acids, e.g. valproic acid having an amino group
    • A61K31/197—Carboxylic acids, e.g. valproic acid having an amino group the amino and the carboxyl groups being attached to the same acyclic carbon chain, e.g. gamma-aminobutyric acid [GABA], beta-alanine, epsilon-aminocaproic acid or pantothenic acid
    • A61K31/198—Alpha-amino acids, e.g. alanine or edetic acid [EDTA]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/34—Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyesters, polyamino acids, polysiloxanes, polyphosphazines, copolymers of polyalkylene glycol or poloxamers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/04—Anorexiants; Antiobesity agents
    • 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
    • A61M37/0015—Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin by using microneedles
    • A61M2037/0023—Drug applicators using microneedles
    • 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
    • A61M37/0015—Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin by using microneedles
    • A61M2037/0046—Solid microneedles
    • 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
    • A61M37/0015—Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin by using microneedles
    • A61M2037/0053—Methods for producing microneedles

Definitions

  • the invention pertains generally to the technical field of medical devices equipped with microneedle (MN) arrays for transdermal administration, and specifically relates to a microneedle patch (MN-patch) loaded with a fat browning agent and a method for preparing the same.
  • MN microneedle
  • MN-patch microneedle patch
  • WAT white adipose tissue
  • BAT brown adipose tissue
  • the hyperplasia and hypertrophy of adipocytes results in a large amount of WAT, and the excess WAT is an important factor leading to diseases, such as type I I diabetes, hypertension, coronary heart disease, stroke, and certain types of cancers (e.g.. colorectal cancer).
  • diseases such as type I I diabetes, hypertension, coronary heart disease, stroke, and certain types of cancers (e.g.. colorectal cancer).
  • the conventional modes of administration such as oral or intravenous injection suffer from problems such as poor absorption, prone to dilution by blood, and degradation by some enzymes in the blood and liver.
  • these conventional modes of administration often result in systemic side-effects or accumulation in non-targeted organs due to systemic administration which requires large doses.
  • Even the FDA-approved drugs, which rely on oral intake, also inevitably cause side- effects.
  • Lorcaserin may cause headache and depression; Orlistat has significant GI side- effects.
  • the drug fen-phen (a combination of appetite suppressants) has a potential to cause pulmonary hypertension and heart valve problems.
  • the drug Sibutramine an appetite suppressant could trigger severe cardiovascular problems.
  • Microneedle (including microneedle array) based transdermal administration is a novel transdermal administration mode.
  • the microneedle first pierces the stratum corneum in the skin by virtue of its sharp micron-scale tip to form a channel to the epidermis or even the upper dermis, which allows the drug to reach the epidermis or upper dermis and then be absorbed by the subcutaneous tissues.
  • Using this method to deliver drugs is not only painless and minimally invasive, but also minizes the toxic side-effects in non-targeted tissues and the first pass effect of the liver, thereby improving the bioavailability.
  • microneedle-based administration can implement self- administration by the patient, which is safe and convenient. Therefore, the microneedle-based transdermal administration technology has gained wide attention from the beginning.
  • microneedle-based transdermal administration technology has drawn more and more attention.
  • silicon microneedles Although the production process of silicon microneedles is mature and they have been widely used, they have poor biocompatibihty with the human body, high price, relatively brittle texture, and complicated production procedures.
  • Metal materials have high mechanical strength and are not easily broken, however, they have the disadvantages of poor biocompatibihty. complicated process and high processing cost, etc.
  • microneedles made of these materials such as metal and silicon are likely to break during insertion into skin.
  • the current research hotspot for the microneedle matrix material is biodegradable polymer materials with good biocompatibility. After the tip of the microneedle is inserted into the skin, the microneedle can be degraded on its own without residues, which solves the problem that the microneedle is difficult to handle once it breaks inside the skin.
  • the matrix material of the biodegradable polymer microneedles should have the following characteristics simultaneously: ( 1) after processing and forming, the material should have sufficient mechanical strength to pierce the stratum corneum, so as to ensure the smooth penetration of the drug loaded in the microneedle; (2) the material is biodegradable without long-term residues; (3) the material has good biocompatibility and does not destroy the inherent structure of proteins in the skin; (4) the material can release the drugs in a desired kinetics, and maintain an effective concentration of the drugs necessary for treatment for a period of time; (5) the material does not affect the biological activity of the drugs; (6) the material is simple and safe to use, even if it is used without medical training, there is no residues of the needle tip or bio-hazardous substances, which will cause no harm to the patient.
  • Maltose, dextran, dextrin, carboxymethyl cellulose, amylopectin, galactose, polylactic acid, polyglycolic acid, chondroitin sulfate, polydioxanone, hyaluronic acid (HA), poly(lactic-co-glycolic acid) (PLGA), etc. can be used as the matrix materials of the biodegradable microneedles.
  • the saccharides have low mechanical strength, and are unstable under high humidity conditions and easily become sticky and deliquescence.
  • Poly lactose and polyglycolic acid have high melting temperatures and require higher temperatures in the preparation of the microneedles. Therefore, they are not suitable for loading of temperature-sensitive drugs.
  • HA non-crosslinked HA, crosslinked HA or HA derivatives
  • hyaluronic acid is an important physiologically active substance in the body.
  • the non-crosslinked HA dissolves and degrades fast, and its half-life is 1-2 days.
  • the dissolution rate and degradation rate of the non-crosslinked HA can be regulated by cross-linking or derivatization.
  • PLGA (or its derivatives), whose backbone is formed by polymerization of two monomers, that is lactic acid and glycolic acid, is a degradable functional polymer organic compound. It is non-toxic and has good biocompatibility, and excellent encapsulation and molding properties. By changing the ratio of lactic acid and glycolic acid in the polymer, dendritic or stellate shape of the polymer, the molecular weight of the polymer, structure derivation, etc. the dissolution and degradation properties of PLGA can be regulated.
  • PLGA has been certified by FDA in the United States and is officially included as a medicinal excipient in the United States Pharmacopoeia.
  • a microneedle patch loaded with a fat browning agent comprising a substrate and an array of microneedles standing on the substrate, wherein the microneedles are made of a polymer material and has a pyramidal structure, the polymer material is a hyaluronic acid or a poly(lactic-co-glycolic acid) or a mixture of both, and the fat browning agent is loaded into the microneedles by pouring into a mold together with the polymer material.
  • a microneedle patch wherein the hyaluronic acid is non-crosslinked, cross-linked or hyaluronic acid derivative.
  • the hyaluronic acid has a molecular weight of less than 10 kDa.
  • the poly(lactic-co-glycolic acid) is a mixture of any two compounds in a mass ratio of 1 : 1 which are selected from the group consisting of a long chain poly (lactic-co-gly colic acid) with a molecular weight of 76,000-1 16,000 Da. a short chain poly(lactic-co-glycolic acid) with a molecular weight of 7,000-17,000 Da. or a star-shaped poly(lactic-co-glycolic acid) with a glucose core having a molecular weight of not more than 15,000 Da: the long chain and short chain poly(lactic-co-giycolic acid) have a structural formula as shown in formula (I):
  • the poly(lactic-co-glycolic acid) is a mixture of poly(lactic-co-glycolic acid) RG756S and poly(lactic-co-glycolic acid) RG 502.
  • the polymer material is a mixture of hyaluronic acid and poly(lactic-co-glycolic acid) in a mass ratio of 1 :99-99: l .
  • the fat browning agent has a mass percentage content of 0.1-50% in each microneedle.
  • the fat browning agent is an adrenergic hormone, thyroid hormone, p3-adrenergic receptor agonist, PPAR agonist, prostaglandin E2 and its analogues, dinitrophenol, fibroblast growth factor 21 , bone morphogenetic protein 7, Apelin, adiponectin or microRNA.
  • the fat browning agent is a ⁇ 3- adrenergic receptor agonist or a thyroid hormone.
  • the p3-adrenergic receptor agonist is Mirabegron or CL316,243.
  • the thyroid hormone is 3,3,5- triiodothyronine or levothyroxine.
  • each microneedle is substantially a quadrangular pyramid in shape, with a needle height of 100-2,000 ⁇ , a height to base width ratio of 2-4, and the microneedle patch have an interval between each microneedle of about 100-2,000 ⁇ .
  • the array of microneedles has one or more microneedle arrays, comprising a 10 ⁇ 10 pyramidal microneedle array, a total of 100 microneedles.
  • the substrate is a sheet made of hyaluronic acid, with a thickness of 1-5 mm.
  • a method for preparing a microneedle patch comprising the steps of dissolving a fat browning agent and a polymer material in a solvent to obtain a molding solution; pouring the molding solution into a cavity of a mold comprising an array of micropores; making the molding solution fill up the micropores; removing any molding solution excess; drying the molding solution to remove the solvent and to form microneedles; filling up the cavity of the mold with a substrate solution containing a polymer material and no fat browning agent; and drying the substrate solution to form a substrate.
  • the polymer material is a hyaluronic acid or a poly(lactic-co-glycolic acid) or a mixture of both
  • the solvent is distilled water, saline, dime thy lformamide or water-dimethylformamide mixture
  • the fat browning agent is an adrenergic hormone, thyroid hormone, p3-adrenergic receptor agonist, PPAR agonist, prostaglandin E2 and its analogues, dinitrophenol, fibroblast growth factor 21 , bone morphogenetic protein 7, Apelin, adiponectin or microRNA.
  • the poly(lactic-co-glycolic acid) is a mixture of poly(lactic-co-glycolic acid) RG756S and poly(lactic-co-glycolic acid) RG 502.
  • the polymer material has a concentration of 0.05-2.0 g/niL.
  • the fat browning agent has a mass ratio to the polymer material of 0.001-0.5: 1.
  • Fig. 1 show's the schematic diagram illustrating transdermal delivery of the drug to the subcutaneous adipose tissue using a dissolvable polymer drug-loaded patch.
  • Fig. 2 shows the characterization of HA microneedle patches.
  • A shows the schematic diagram illustrating the preparation processes of the Cy5-loaded and drug-loaded microneedle patches.
  • [B] shows the photograph of an 11 A microneedle patch.
  • D shows the diagram of acting force vs displacement of the unloaded and drug-loaded microneedle patches.
  • El] and [E2] show the SEM photographs of an HA microneedle before and after mechanical loading respectively. Scale - 100 ⁇ .
  • Fig. 3 shows the in vitro and in vivo dissolution and release of drug-loaded HA microneedles.
  • A shows the in vitro release profiles of Cy5 from a Cy5-loaded HA microneedle patch in phosphate buffer (PBS, pH 7.4) and in mouse serum.
  • B shows the release profile of CL316,243 from a CL316,243-loaded HA microneedle patch in porcine skin.
  • FIG. 4 shows the in vivo dissolution and release of drug-loaded HA microneedles.
  • C I] and [C2] show the photographs of porcine skin after application and removal of a CL316,243-loaded HA microneedle patch, respectively. Scale- 2 mm.
  • [Fl] and [F2] show the microscope photographs of skin sections before and after administration of a microneedle patch, respectively.
  • Fig. 5 shows the in vivo distribution of drug-loaded HA microneedles.
  • G] and [H] show the in vivo fluorescence photographs of Cy5 after administration for 2 hours and 18 hours for the control and CL316,243-loaded microneedles, respectively.
  • I] shows the fluorescent photographs of the organs and subcutaneous adipose tissue (Inguinal WAT) treated with a Cy5-loaded microneedle patch for 2 hours and 18 hours.
  • [Jl] and [J2] show the fluorescent intensities of the organs and subcutaneous adipose tissue (Inguinal WAT) in mice after intraperitoneal injection of Cy5 and administration of Cy5 -loaded microneedles.
  • Fig. 6 shows the schematic diagram of administration sites in mice. [A2]-[A4] show the photographs of administration mode.
  • FIG. 7 shows the in vivo experiment of drug-loaded HA microneedle patches for obesity treatment. Mice were treated for 5 consecutive days. An unloaded microneedle patch was applied for the control, and a drug-loaded microneedle patch was applied for the experimental group.
  • [E] shows the photographs of total inguinal WAT, epididymal WAT and interscapular BAT in mice treated with different methods.
  • [F] and [G] show the ratio of adipose tissue weight to body weight after treatments by different methods.
  • Fig. 8 shows the expression of WAT browning markers in groins of mice treated differently.
  • [H] shows the results of Western blots.
  • [J1]-[J4] show the Western blotting analyses of UCP1, COX1, PGCla and aP2.
  • Fig. 9 shows the anti-obesity effects of CL316,243 administered by transdermal and injection routes.
  • [A] shows the photographs of total inguinal WAT, epididymal WAT and interscapular BAT in mice after injection with 2.5 g CL316,243 or 0.5 ⁇ g T3 for 5 days.
  • [B l] shows the body weights, and [B2] shows the ratio of adipose tissue weight to body weight.
  • Fig. 10 shows the comparison of mice injected with phosphate buffer or high dose of CL316,243 (12.5 ⁇ g) for 5 consecutive days or administered transdermally with a CL316,243 -loaded microneedle patch (total 100 microneedles, about 2.5 ⁇ g drug).
  • C shows the schematic diagram of administration sites.
  • D1]-[D2] show the body weights and ratio of adipose tissue weight to body weight.
  • [E] shows the photographs of total inguinal WAT. epididymal WAT and interscapular BAT.
  • Fig. 11 shows the diagram of body surface temperature changes.
  • [F] shows the result of infrared thermal imaging analysis.
  • [G] shows the comparison of the temperatures of control (without drug), high-dose injection group and low-dose microneedle patch group.
  • Fig. 12 shows the photographs of the Hematoxylin and Eosin (H&E) stained inguinal WAT in mice treated differently.
  • H&E Hematoxylin and Eosin
  • Fig. 14 shows the characterization of PLGA microneedle patches.
  • A shows the schematic diagrams of the drug delivery systems based on a long-chain PLGA and a branched PLGA with a glucose core.
  • B shows the bright-field microscope photographs and confocal microscope photographs of Cy5-loaded PLGA microneedle patches.
  • C shows the in vitro release profiles of Cy5 from Cy5-loaded PLGA microneedle patches in phosphate buffer (PBS. pi I 7.4).
  • PBS phosphate buffer
  • D shows the mechanical properties of the Cy5-loadcd PLGA microneedle patches.
  • [E] and [F] show the SEM photographs of the CL316,243-loaded PLGA microneedle patches before and after insertion into the porcine skin, respectively.
  • [G] and [H] show the photograph of the porcine skin inserted by one microneedle and the bright-field photograph of the stained porcine skin with insertion marks.
  • Fig. 15 shows the in vivo anti-obesity experiment of CL316,243-loaded PLGA microneedle patches. Mice were dosed twice a week and observed for 25 days.
  • [A] shows the photographs of the sites of hair removal treated with microneedles for 0 day and 5 days.
  • [B] shows the fluorescent photographs of the mice treated with microneedle patches for 0 day and 5 days.
  • [C] shows the average body weight increase profiles of the mice dosed differently.
  • [D] shows the average daily feed intake in mice.
  • E] and [F] show the photograph of total inguinal WAT, epididymal WAT and interscapular W T AT and the diagram of the ratio of adipose tissue weight to body weight.
  • [G] shows the result of infrared thermal imaging analysis.
  • [H] shows the diagram of body surface temperature changes.
  • Fig. 16 shows the results of WAT browning by CL316.243-loaded PLGA microneedle patches in diet-induced obese mice.
  • Al] and [A2] show the photographs of the stained inguinal WAT in mice dosed differently.
  • B5 ] show the Western blotting analyses of UCP1, COXl . PGCla, and aP2.
  • C]-[G] show the quantitative results of cholesterol, free fatty acids, triglycerides, glucose and insulin in the serum from mice dosed differently.
  • FIG. 17 [A] shows the in vivo pharmacokinetics profiles of T3 in mice treated with a ⁇ 3 -loaded microneedle patch and injected with T3. [B] shows the average weights of mice treated differently. [C] and [D] show the levels of T3 and TSH in the serum of the T3-treated and control mice.
  • the invention provides a microneedle patch loaded with a fat browning agent.
  • a fat browning agent By loading the fat browning agent into a polymeric microneedle array integrated onto a disposable patch for transdermal administration, direct and local drug delivery to the subcutaneous white adipose tissue (WAT) can be achieved, which promotes browning of WAT effectively, and thus suppresses fat and weight gain without daily dietary control.
  • WAT subcutaneous white adipose tissue
  • the invention provides a microneedle patch loaded with a fat browning agent, comprising a substrate and an array of microneedles standing on the substrate, wherein the microneedle is made of a polymer material and has a pyramidal structure, the polymer material is hyaluronic acid, poly(lactic- co-glycolic acid) or a mixture of both, and the fat browning agent is loaded into the microneedles by pouring into a mold together with the polymer material.
  • the white fat tissue of the human body is mainly distributed in the abdomen and groin.
  • This invention chooses suitable degradable polymers according to the characteristics of the drug application locations.
  • Hyaluronic acid (HA) is a linear macromolecular acid mucopolysaccharide widely distributed in the extracellular matrix of the soft connective tissues, which is comprised by disaccharide units of glucuronic acid and N-acetyl glucosamine alternately connected together, and is an important constitute of skin, vitreous body and cartilage tissue. Therefore, hyaluronic acid is used to prepare the microneedles of the invention.
  • the hyaluronic acid is a non-crosslinked hyaluronic acid, crosslinked hyaluronic acid or hyaluronic acid derivative.
  • the molecular weight of the hyaluronic acid is less than 10 kDa.
  • the non-crosslinked hyaluronic acid is not suitable for applications requiring slow release due to its high dissolution rate and degradation rate, and can be combined with a drug having a short half-life to achieve rapid drug release.
  • the crosslinked hyaluronic acid or hyaluronic acid derivative can be used in the applications requiring slow drug release.
  • the degradation products of poly(lactic-co-glycolic acid) are lactic acid and glycolic acid. They are the by-products of human metabolic pathway, and thus have no toxic side-effects to human body. Therefore, they are suitable for use in the invention.
  • PLGA has been certified by FDA and has been officially included as a medicinal excipient in the United States Pharmacopoeia.
  • the dissolution rate of PLGA is slow and it can be combined with a drug having a long half-life.
  • the solubility of PLGA can also be regulated by adjusting the molecular weight, alteration of branching or star- shaped structure, or derivatization.
  • the PLGA raw material used includes a long-chain PLGA with a molecular weight of 76,000- 1 16,000 Da, a short chain PLGA with a molecular weight of 7,000-17,000 Da or a star-shaped PLGA with a glucose core having a molecular weight of not more than 15,000 Da.
  • the long chain and short chain PLGAs have a structural formula as shown in formula (I):
  • the star-shaped PLGA with a glucose core has a structural formula as shown in formula (II):
  • the drug release kinetics and microneedle mechanical properties can be readily regulated by the mixture of different types of PLGA or their derivatives as well as the component ratio.
  • the PLGA is a mixture of any two compounds in a mass ratio of 1 : 1 which are selected from the group consisting of a long chain PLGA, a short chain PLGA, or a star-shaped PLGA.
  • the polymer material can be a mixture of HA and PLGA in a mass ratio of 1 :99- 99: 1.
  • the drugs for preventing or treating obesity is classified into appetite suppressants, digestive adsorption blockers, hypoglycemic agents (such as biguanides, a-glucosidase inhibitors, GLP-1 and DPP-4 inhibitors) and fat browning agents.
  • the fat browning agents are a class of metabolic stimulators, which can promote browning of WAT, whereby boosting the thermogenesis process of the brown adipose tissue and the decomposition and oxidation of fat and glycogen in the body. In comparison with other anti-obesity drugs, weight and body fat can be reduced without affecting the food intake.
  • the fat browning agents used in the invention include adrenergic hormone, thyroid hormone, ⁇ 3 -adrenergic receptor agonist, PPAR agonist (such as PPARy agonist), prostaglandin E2 and its analogues, dinitrophenol, fibroblast growth factor 21, bone morphogenetic protein 7, Apelin, adiponectin, microRNAs and the like.
  • the fat browning agent is a ⁇ 3 -adrenergic receptor agonist or a thyroid hormone.
  • the p3-adrenergic receptor is Mirabegron.
  • the p3-adrenergic receptor is CL316,243.
  • the thyroid hormone is 3,3,5-triiodothyronine.
  • the thyroid hormone is levothyroxine.
  • the microneedle array can realize the direct and local drug delivery to the subcutaneous WAT, the required concentration of the drug is low, but the bioavailability is high. As a result, the cost and possible side-effects can be reduced while the efficacy is ensured and the concentration of the drug can be as low as possible.
  • the fat browning agent has a mass percentage content of 0.1-50% in each microneedle.
  • the drug is uniformly distributed in the array of microneedles.
  • the polymer microneedles of the invention in which the drug is uniformly distributed have high drug loading.
  • the human skin has three layers of tissue: stratum corneum, active epidermis and dermis.
  • stratum corneum the thickness of the outermost stratum corneum is 10-40 ⁇ , and it is composed of dense keratinocytes and hence is the major obstacle to drug delivery.
  • the thickness of the epidermis is 50-100 ⁇ , and it contains active cells and a small amount of nerve tissue, but does not contain blood vessels.
  • the dermis is the main part of the skin, containing numerous viable cells, a large amount of nerve tissue and blood vessel tissue, and its thickness is about 2,000 ⁇ .
  • the microneedles In order to guarantee the high permeability and avoid pain during the piercing of microneedles, the microneedles should be able to pierce the stratum corneum of the skin to reach the epidermis and enter part of the dermis. Therefore, the total number of microneedles in the microneedle array, the height of the microneedle, the interval between microneedles and the base width of the microneedle need to be considered comprehensively in order to achieve piercing into the skin by thumb pressing and generate as little pain as possible.
  • the microneedle is substantially a quadrangular pyramid in shape, with a microneedle height of 100-2,000 ⁇ , a height to base width ratio of 2-4, and an interval between microneedles of about 100-2,000 ⁇ .
  • the microneedle is a quadrangular pyramid in shape, with a microneedle height of 100-900 ⁇ , a height to base width ratio of 2-2.5, and an interval between microneedles of 100-900 ⁇ . It is found by careful selection that the pyramid-based pyramidal microneedle array with a height to base width ratio of 2. and an interval between microneedles of 700 ⁇ archives the best effect of penetration.
  • the array of microneedles can be in a rectangular, quadrangular, circular or oval shape.
  • the array of microneedles has one or more microneedle arrays, comprising a 10 ⁇ 10 pyramidal microneedle array and a total of 100 microneedles. More preferably, 10 x 10 pyramidal array with a microneedle height of 600 ⁇ and a microneedle base width of 300 ⁇ is employed.
  • the function of the substrate is to provide support for the microneedle array, so that the skin can be pierced simply by pressing.
  • the therapeutic drug molecules are only loaded in the microneedles but not the substrate, without requiring a large area of supporting substrate, which effectively reduce the waste of expensive drugs.
  • the substrate is made of the same or different polymer material as the microneedles.
  • the substrate is a sheet made of HA with a thickness of 1-1.5 mm.
  • HA has good biocompatibility and low price. Therefore, after the microneedle is inserted into the skin, the microneedles can be detached from the substrate automatically due to the moisture on the surface of the body or the subcutaneous body fluid inhaled by the microneedle.
  • the microneedle patch of the invention can be applied as follows: the patch with an array of disposable drug-loaded polymer microneedles loaded with a fat browning agent is applied on the skin, the polymer microneedles are pierced into the skin by pressing, and the drug is released after the polymer microneedles are dissolved, to achieve the effect of preventing or treating obesity. After the microneedles are pierced, the substrate patch can be removed directly.
  • the invention provides a method for preparing a microneedle patch, comprising the steps of: dissolving a fat browning agent and a polymer material in a solvent to obtain a molding solution;
  • a polydimethylsiloxane (PDMS) microporous mold is used in the invention to prepare the drug-loaded polymer microneedles.
  • the PDMS microporous mold is prepared by mixing PDMS and a curing agent methyl dimethicone in a mass ratio of 10: 1 and pouring the mixture into a stainless steel mold, and performing injection molding, followed by degassing in vacuum and curing at 70 °C for 2-5 hrs.
  • the stainless steel mold contains one or more female molds of pyramidal microneedle array.
  • the stainless steel mold comprises one or more 10 ⁇ 10 pyramidal arrays, each array comprising 100 microneedles and interval between microneedles of 700 ⁇ , each microneedle has a height of 600 ⁇ , and a microneedle base width of 300 ⁇ .
  • the mixture of the fat browning agent and HA or PLGA or the HA-PLGA mixture is added into the PDMS micromold, then centrifugation is conducted and drying is performed at room temperature for a certain period of time.
  • the preparation process does not require such procedures which are prone to destroy the structure of microneedles, the composition and activity of the drugs (e.g., UV irradiation and heating).
  • the polymer material used in the method is a hyaluronic acid or a poly(lactic-co-glycolic acid) or a mixture of both.
  • the solvent used in the method is distilled water, dimethylformamide or water- dimethylformamide mixture.
  • the fat browning agent used in the method is an adrenergic hormone, thyroid hormone, p3-adrenergic receptor agonist, PPAR agonist, prostaglandin E2 and its analogues, dinitrophenol, fibroblast growth factor 21, bone morphogenetic protein 7, Apelin, adiponectin or microRNA.
  • the invention employs the corresponding solvents, drugs and proportion ratios according to the characteristics of the polymer materials.
  • the solvent for HA is distilled water and the concentration of HA is 0.05-2.0 g/mL.
  • HA has good molding property, however, when the concentration of HA is too low, the mechanical strength of the resultant microneedle is low, and when the concentration of HA too high, the resultant microneedle is prone to absorb moisture.
  • the preferred concentration of HA is 0.5 g/mL. Studies have shown that the mechanical force of the microneedle prepared at this concentration is 0.06 N per needle, which is sufficient to pierce skin and have the ability to resist compression.
  • the mass ratio of the drug to HA is 0.001-0.5: 1, preferably 0.005: 1 (e.g., 2.5 mg: 0.5 g).
  • the solvent for PLGA is dimethylformamide.
  • the concentration of PLGA is 0.05-1.0 g/mL, preferably 0.2 g/mL; the mass ratio of the drug to PLGA is 0.01-0.5: 1, preferably 0.05: 1 (e.g., 10 mg: 200 mg).
  • the solvent is a water-dimcthylformamide mixture.
  • the percentage content of dimethylformamide is 1-99%, and the mass ratio of the drug to the HA-PLGA mixture is 0.001-0.05: 1 .
  • the aforementioned two polymer materials not only have good molding properties, but also have good compatibility with the fat browning agents and do not affect the normal efficacy of the fat browning agents.
  • the microneedle-based transdermal administration in the invention only needs 1/5 of the drug used in the intraperitoneal injection to achieve the same anti-obesity effect. It is proved that the compatibility of the two polymers with the fat browning agent is good while the normal efficacy of the fat browning agent is not affected.
  • centrifugation is adopted to fill the polymer solution into the PDMS mold quickly, and to promote the uniform distribution of the drug.
  • the centrifugation rate and time are mainly considered from the perspectives of energy consumption and easy operation.
  • the centrifugation is performed at a rate of 1 ,000-20,000 rpm, preferably 4,000 rpm, for 1-20 min, preferably 3 min. Drying is performed at room temperature for 4-18 hrs, preferably 12 hrs.
  • the substrate solution comprising a polymer material at a concentration of 0.05- 2.0 g/mL is added at the bottom of the microneedle array to form the substrate.
  • the solution of HA 0.5 g/ml
  • secondary centrifugation 4,000 rpm for 3 min
  • drying 12 hrs at room temperature
  • the microneedles are peeled off from the mold and stored at 4 °C.
  • the microneedles are perpendicular to the patch substrate, and preferably the patch substrate has a size of approximately 6 mm x 6 mm.
  • the beneficial effects of the invention are as follows: ( 1 )
  • the microneedle patch with an array of microneedles loaded with a fat browning agent provided by the invention can directly deliver the drug subcutaneously to the subcutaneous adipose tissue.
  • the drug-loaded microneedles can be embedded into the skin for administration after brief thumb pressing. Because the microneedles do not need to be pulled out, skin discomfort and irritation are avoided.
  • patients can use the microneedle patch conveniently and effectively, thereby achieving obesity control or treatment of other metabolic diseases at home.
  • HA and PLGA have good compatibility with the fat browning agents. These polymers, together with the centrifugation operation during the preparation procedures, ensure uniform distribution and high loading of the drug in the microneedle array. In addition, the drug release kinetics can be regulated by adjusting the composition of the polymer materials.
  • microneedle-based transdermal administration in the invention does not alter the chemical structure and efficacy of the drug itself.
  • the microneedle patch provided by the invention has the significant advantages of low-dose administration, and local targeted administration, avoiding the issues such as drug failure or severe side-effects caused by conventional systemic administration.
  • the patch with an array of microneedles in the invention the gain of WAT and body weight can be suppressed without daily dietary control, and obesity can be prevented or treated.
  • This invention for the first time realizes direct delivery of fat browning agents to adipose tissues, which can prevent obesity, reduce weight and improve the levels of metabolites in the serum such as total cholesterol, free fatty acids, insulin, glucose and triglycerides.
  • the invention is suitable for home-based long-term self-actualized prevention, management or treatment of obesity and associated diseases.
  • mice used were C57BL/6J mice, hyaluronic acid (HA), CL316,243, T3, long-chain PLGA (PLGA RG756S, poly(D,L-lactic-co-glycolic acid), 75:25. molecular weight 76,000-1 16,000), short-chain PLGA (PLGA RG502, poly(D,L-lactic-co-glycolic acid), 50:50, molecular weight 7,000-17,000), star-shaped PLGA with a glucose core (PLGA 5 arm star) 15,000 purchased from Sigma-Aldrich.
  • the application for preventing or treating obesity was confirmed by mouse experiments. All data were analyzed using t-test and the results were expressed as average values.
  • adipose tissue was based on total inguinal white adipose tissue (IgWAT), epididymal white adipose tissue (EpiWAT) and interscapular brown adipose tissue (BAT).
  • the PDMS micromold was fabricated by pouring PDMS (the mass ratio of PDMS to the curing agent methyl dimethicone was 10: 1) into a stainless steel mold (10 x 10 pyramidal array, total 100 microneedles, microneedle height of 600 ⁇ , interval between microneedles of 700 ⁇ , microneedle base width of 300 ⁇ ), followed by degassing in vacuum and curing at 70 °C for 2 hours.
  • EXAMPLE 1 The polymer selected was HA. and the active fat browning agent selected was ⁇ 3 -adrenergic receptor agonist CL316,243.
  • the administration mode of the microneedle patch is as shown in Fig. 1.
  • Fig. 2A Preparation of the microneedles is as shown in Fig. 2A.
  • a drug-containing HA solution 0.5 g of HA was dissolved into 1 ml of distilled water, and then 2.5 mg of CL316,243 was added
  • the patch substrate was made by further preparing an HA layer at the bottom of the microneedle array.
  • An HA solution (0.5 g/ml) was added at the bottom of the microneedle array. After secondary centrifugation (4,000 rpm, 3 min) and drying at room temperature for 12 hours, the microneedles were peeled off from the mold and then stored at 4 .
  • the photograph of the prepared microneedles is as shown in Fig. 213.
  • Fig. 2 (C1-C3) The fluorescence micrograph photographs of a microneedle patch loaded with both a drug and fluorescent dye Cy5 are as shown in Fig. 2 (C1-C3). It can be seen that Cy5 was uniformly distributed in the microneedles.
  • the characterization of the microneedle mechanical properties is as shown in Fig. 2D, which indicates that each microneedle can withstand more force than that required for piercing the skin. This result ensures that the microneedle can pierce the skin simply by thumb pressing.
  • Fig. 2 (El) is a scanning electron microscopy (SEM) photograph of the microneedles before the mechanical property test. It can be seen that the microneedle array has a pyramidal structure.
  • E2 is an SEM photograph of the microneedles after the insertion of microneedles and removal of the patch substrate, which shows the fracture at the root of the microneedles.
  • the mechanical properties of the microneedle can ensure that it can pierce into the skin >350 ⁇ without breaking.
  • the disposable drug-loaded patch with a polymer microneedle array was affixed to the skin of the mice. After the polymer microneedles were dissolved, the drug was released to achieve the prevention or treatment of obesity.
  • the 3-adrenergic receptor agonist CL316,243 can promote the lipolysis of the white adipose tissue (WAT), stimulate the non-trembling heat production in the brown adipose tissue, inhibit the differentiation of adipocytes, and induce the appearance of brown adipocytes in WAT. As a result, it can consume WAT to achieve anti-obesity effect.
  • WAT white adipose tissue
  • the effect of preventing or treating obesity was demonstrated by mouse experimental models with diet-induced obesity. Compared with the control, the mice after subcutaneous administration exhibited reduced adipose tissue mass.
  • the changes of serological related parameters are used as an auxiliary evaluation method.
  • Cy5 molecules can be rapidly released in PBS with a half- life ( ⁇ ) of ⁇ 3.5 seconds due to the rapid dissolution of HA. It also can be rapidly released in the mouse serum with a half-life ⁇ of ⁇ 5.6 seconds.
  • the amount of the drug that enters the skin was determined by subtracting the amount of the residual drug in the microneedles from the initial amount of the drug.
  • the amount of the residual drug in the microneedles was obtained by pulling out the microneedles and then dissolving them in PBS (pH 7.4); the amount of the drug that remained in the skin could also be obtained by soaking the punctured skin in PBS.
  • the extracted CL316,243 was analyzed using reversed-phase liquid chromatography.
  • the chromatographic column was Agilent Poro shell 120 EC-C18 with a mobile phase of water: methanol (at a volume ratio of 80:20), a flow rate of 0.5 mL/min. UV detection was performed at 230 nm, and the external standard method is adopted for quantification.
  • Fig. 4 (C1-C2) shows the result of the skin piercement test, which demonstrates that the HA microneedle can be rapidly dissolved in 2 minutes without obvious skin erythema, swelling and infection.
  • Fig. 4D-E show the SEM photographs of the microneedles before and after transdermal administration. After 2 minutes of applying HA microneedles loaded with CL316-243, the microneedles were embedded into the skin well and completely dissolved. Obviously, the therapeutic agent can be quickly and fully delivered into the skin by the microneedles of the invention.
  • mice were affixed to the left or right groin of the mice. After 5 minutes, the patches were removed and the mice were euthanized and dissected. The distribution of Cy5 in WAT and other viscera at the subcutaneous insertion site of microneedles was examined. In order to study the difference in the efficacy between subcutaneous microneedle administration and intraperitoneal injection, the mice were administered by microneedles in the experimental group, while the mice were mtraperitoneally injected with Cy5 (1 ⁇ g in 50 ⁇ PBS) in the control experiment. The subcutaneous WAT and the main viscera such as liver, heart, kidney, and lung obtained from the dissected mice in the experimental and control groups were used for fluorescence imaging.
  • Fig. 5G The result is shown in Fig. 5G.
  • Fig. 51 and Fig. 5(J1 -J2) when the microneedle was administered for 2 hours and 18 hours, no fluorescence signal was detected in organs such as the heart, head and neck, and there was only the background auto-fluorescence produced from food in stomach and intestine.
  • mice injected with Cy5 is in stark contrast to that of mice with the subcutaneous microneedle administration in present invention.
  • Most of the Cy5 was distributed in liver and kidney, resulting in hepatic metabolism and renal clearance. Therefore, local diffusion and transdermal drug molecule accumulation can be regarded as an effective method for targeting subcutaneous WAT for the treatment of obesity.
  • mice experiments to test the anti-obesity effect of drug-loaded microneedles
  • C57BL/6J mice (7-8 weeks old, male) were normally fed (12 hours in light/12 hours in darkness, grown at 21°C), and water and diet were provided following the normal standards during feeding.
  • mice were randomly divided into three groups.
  • the microneedle patch used was a drug-loaded microneedle patch (the microneedles in the patch were loaded with ⁇ 2.5 ⁇ g of CL316,243 respectively).
  • the daily dosage for microneedle patch administration was 2.5 ⁇ g CL316,243 (-0.1 mg/kg/day). Drugs were all dissolved in 100 ⁇ PBS.
  • the amount of CL316,243 loaded in the microneedles was accurately determined by HPLC or enzyme-linked immunosorbent assay (ELISA).
  • the unloaded patch was used as control for testing the effect of the drugs.
  • the mice were mtraperitoneally injected with a high dose of CL316,243 everyday (0.5 mg/kg/day) to compare the delivery efficiencies of drugs by transdermal administration or intraperitoneal injection.
  • mice were subcutaneously administered drugs daily for 5 consecutive days.
  • the microneedle administration method is as shown in Fig. 6 (A1-A4).
  • the mice hair in the inguinal region (lower left or right dorsolateral area, close to the hindlimb) was carefully shaved, and the microneedle patch was placed on the left or right side of the exposed area and remained for 5 minutes to allow the microneedles to completely dissolve.
  • the body surface temperature in differently treated mice was measured using an infrared thermography to reflect the body temperature of the mice.
  • mice Five days later, lethal doses of carbon dioxide were provided to euthanize the mice. After body weight was measured, the inguinal WAT (IgWAT), epididymal WAT, and interscapular BAT were individually cut out and weighed.
  • IgWAT inguinal WAT
  • epididymal WAT epididymal WAT
  • interscapular BAT were individually cut out and weighed.
  • Fig. 7E and Fig. 7F show that the white fat on the left and right sides of the groin of the mice was significantly reduced in the drug group (0.1 mg/kg/day). Although the administration of microneedles was performed only on either left side or right side of the groin, the volume of the white fat at both sides of the groin was reduced compared with the control mice administered with no drug. In addition, CL316,243-treated mice exhibited significantly reduced epididymal WAT (peritoneal visceral fat), suggesting that the transdermal release of CL316,243 has a systemic anti-obesity effect (Fig. 7E and Fig. 7F). Compared with the microneedle patch loaded with CL316,243, the drug patch loaded with T3 can only reduce the white fat in the administered site, but not in the non-administered site (Fig. 7E-G).
  • Each sample was electrophoresed on 12% SDS-PAGE and then transferred to a nitrocellulose membrane.
  • the membrane was blocked with a blocking buffer for 2 hours at room temperature and then was incubated with a primary antibody (1 :200-400 dilution) at room temperature for 12 hours allowing for specific binding and washed with a Tween-containing Tris buffer (TBST) for 3 x 15 minutes. Thereafter, the membrane was incubated with a horseradish peroxidase-labeled secondary antibody (1 :2,000-4,000 dilution) at room temperature for 6 hours and then was washed with TBST for 3 x 15 minutes. Protein bands were detected with an imaging system.
  • the antibodies used were UCP1 (sc-6529), aP2 (sc-18661), PGC- ⁇ (sc-13067), COX1 (sc-23982), and actin (sc-1616).
  • the anti-UCPl antibody is PA1-24894.
  • intraperitoneal injection of the same dose of CL316,243 (0.1 mg/kg/day) does not significantly alter the weight of the adipose tissue. Only when the intraperitoneal injection dose is increased by 5 times, the intraperitoneal injection can achieve the same effect as low-dose subcutaneous administration by microneedles.
  • Fig. 1 1 F and Fig. 1 1 G show that both intraperitoneal injection and low- dose microneedle transdermal administration lead to an increase in the body surface temperature, at least in part due to the conversion of adipose tissue into the energy-consuming state (browning) (as shown in Fig. 12).
  • EXAMPLE 2 The polymer selected was PLGA, and the active fat browning agent selected was the p3-adrenergic receptor agonist CL316.243.
  • the patch substrate was made by further preparing an HA layer at the bottom of the microneedle array.
  • An HA solution (0.5 g/ml) was added at the bottom of the microneedle array. After secondary centrifugation (4.000 rpm, 3 min) and diying at room temperature for 12 hours, the microneedles were peeled off from the mold and then stored at 4°C.
  • the disposable drug-loaded patch with a polymer microneedle array was affixed to the skin of the mice. After the polymer microneedles were dissolved, the drug was released to achieve the prevention or treatment of obesity. Immediately after the microneedle patch was inserted, the HA substrate was separated and removed to prevent the production of an HA solution due to rapid dissolution.
  • PLGA Poly(lactic-co-glycolic acid)
  • the microneedle made with the long-chain PLGA has a sharp pyramidal structure while the short-chain and star-shaped PLGA have a non-ideal morphology.
  • the dissolution of the long- chain PLGA and the release of Cy5 molecules are very slow. After being soaked in phosphate buffer for 7 days, the long-chain PLGA could still maintain a good pyramidal structure, and after it was soaked in PBS for 4 days or 2 weeks, only 8.6% or 37.3% of Cy5 molecules were released.
  • the long-chain PLGA. short- chain PLGA or star-shaped PLGA can be mixed. We found that the mixture of the long-chain PLGA and short-chain PLGA at a ratio of 1 : 1 is most desirable. As shown in Fig. l4C-D, the microneedles prepared with PLGA756s+502 are very sharp and can release 31.8% of Cy5 molecules in 4 days, similar to the release effect of the pure short-chain PLGA microneedles or star-shaped PLGA microneedles.
  • the pure long-chain PLGA microneedles have the highest mechanical strength, but the mechanical strength of the microneedles prepared with PLGA756s+502 exceeds the force necessary to pierce the porcine skin. Therefore, PLGA756s+502 is preferred for the preparation of microneedles.
  • Fig. 14E SEM characterization shows that the base width of the microneedle is 250 ⁇ and the height of the microneedle is 570 ⁇ .
  • Fig. 14F after being wetted with PBS rapidly ( ⁇ 1 min), the microneedles were peeled off from the HA substrate. As shown in Fig. 14G and Fig. 14H, these microneedles can be pierced into the porcine skin by thumb pressing, with a penetration depth of -300 um. Slow dissolution properties and appropriate mechanical strength make the drug-loaded PLGA microneedle suitable as a subcutaneous delivery system for sustained drug release.
  • the material for preparing PLGA microneedles was 756s+502. Using an experimental method similar to that for HA microneedle patches, PLGA microneedle patches were also applied to the inguinal region of mice. The microneedles left markers after piercing into the skin, but did not cause any obvious skin abnormalities such as skin erythema, swelling, and infection, as shown in Fig. 15A.
  • Fig. 15B shows that after 5 days of applying PLGA microneedle patches loaded with Cy5, a significant fluorescence signal was observed near the application site, which further confirms the slow dissolution of PLGA. Moreover, such retention time of PLGA microneedles is comparable to the mouse epidermal metabolic turnover time (8-10 days). Obviously, the implanted PLGA microneedles can serve as a long-lasting slow-release reservoir of drugs.
  • mice were C57BL/6J mice (9-10 weeks old, male, weighing about 30 grams). The mice were randomly divided into 3 groups and treated with the drug twice a week. The control group was treated with the unloaded microneedle patches.
  • sample groups were treated with microneedle patches loaded with CL316,243 (each mouse was treated with one patch, and 10 ⁇ g CL316,243 was loaded in microneedles of each patch) or with intraperitoneal injection of CL316.243 (10 ⁇ g per mouse; -0.3 mg/kg).
  • the microneedle patches were used twice a week, and compared with the control mice treated with unloaded microneedles, the CL316,243 -loaded microneedles could significantly reduce -15% of weight gain. In contrast, intraperitoneal injection of the same dose ( ⁇ 0.3 mg/kg, twice weekly) did not significantly prevent weight gain.
  • the brown adipocytes at the inguinal WAT were increased, and all of the browning markers were significantly increased, as shown in FIG. 16 B 1 -135 ). These results prove that browning of WAT is achieved.
  • Sustained transdermal administration is superior to systemic administration.
  • obese mice exhibit metabolic syndromes, including high levels of total cholesterol, free fatty acids, insulin, glucose, and triglycerides, which are high risk factors for a range of metabolic diseases such as diabetes.
  • the microneedle treatment in the invention can improve the levels of the above metabolites in the serum (Fig. 16C-G). Therefore, the microneedle patches in the invention are suitable for the prevention, management or treatment of obesity' and related diseases based on long-term self-administration at home.
  • EXAMPLE 3 The polymer selected was HA, and the active fat browning agent selected was thyroid hormone T3.
  • a drug-containing HA solution (0.5 g of HA was dissolved into 1 ml of distilled water and then 0.5 mg of T3 was added) was added to a PDMS micromold, followed by centrifugation (4,000 rpm, 3 min) and drying at room temperature for 12 hours.
  • the patch substrate was made by further preparing an HA layer at the bottom of the microneedle array.
  • An HA solution (0.5 g/ml) was added at the bottom of the microneedle array. After secondary centrifugation (4,000 rpm, 3 min) and drying at room temperature for 12 hours, the microneedles were peeled off from the mold and then stored at 4°C. [00151] 2. Application in prevention or treatment of obesity
  • the disposable drug-loaded patch with a polymer microneedle array was affixed to the skin of the mice. After the polymer microneedles were dissolved, the drug was released to achieve the prevention or treatment of obesity.
  • TSH thyroid stimulating hormone
  • the levels of free fatty acids, triglycerides and glucose in the serum of obese mice were determined by the standard assay kits (Biovision).
  • the levels of cholesterol and insulin were measured by the cholesterol quantitation kit (Sigma-Aldrich) and mouse insulin ELISA kit (ThermoScientific).
  • the subcutaneous WAT at the transdermally administered site of the mice treated with T3-loaded HA microneedles was lighter and smaller than that of the control mice, and also there was no significant difference in epididymal WAT (visceral fat).
  • the polymer material of the invention does not change the chemical structure and efficacy of the drug itself, and no other auxiliary materials are added during the preparation of microneedles, thereby avoiding the influence of auxiliary materials on the shape and efficacy of the drug (e.g., through electrostatic interaction, hydrophobic interaction, and inclusion effect).
  • FIG. 17B, FIG. 17C, and FIG. 17D show that there is no significant difference in body weight, serum T3 and serum TSH between the control mice and the microneedle- administered mice.
  • Normal serum levels of free T3 and TSH indicate that transdermally-delivered T3 is largely confined near the subcutaneous white adipocytes, because T3 molecules reaching circulation are either bound to the T3 -binding proteins (e.g., globulin, albumin, etc.) or degraded in the liver.
  • Intraperitoneal injection of the same dose of T3 had no effect on the weight of epididymal WAT.
  • the above examples fully illustrate the invention.
  • the polymer species are carefully chosen for the invention.
  • HA and PI G A have good compatibility with the fat browning agents.
  • These polymers together with the centrifugation operation during the preparation procedures, ensure uniform distribution and high loading of the drug in the microneedle array.
  • the drug release kinetics can be regulated by adjusting the composition of the polymer materials.
  • the micronecdlc- based transdermal administration in the invention does not alter the chemical structure and efficacy of the drug itself. Direct administration to subcutaneous adipose tissue can be achieved by microneedle- based transdermal administration.
  • the microneedle patch provided by the invention has the significant advantages of low-dose administration, and local targeted administration, avoiding the issues such as drug failure or severe side-effects caused by conventional systemic administration.
  • the patch with an array of microneedles in the invention the gain of WAT and body weight can be suppressed without daily dietary control, and obesity can be prevented or treated.

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Abstract

L'invention concerne un patch à micro-aiguilles chargé d'un agent de brunissement de graisse et son procédé de préparation, relevant du domaine technique de l'administration par voie transdermique. Le patch à micro-aiguilles comprend un substrat et une matrice de micro-aiguilles agencée verticalement sur le substrat. La micro-aiguille est constituée d'un matériau polymère et a une structure pyramidale. Le matériau polymère est l'acide hyaluronique, un poly(acide lactique-co-glycolique) ou un mélange des deux. L'agent de brunissement de graisse est chargé dans les micro-aiguilles par versement dans un moule conjointement avec le matériau polymère. Les patchs à micro-aiguilles selon l'invention peuvent être directement administrés par voie sous-cutanée au tissu adipeux sous-cutané. Après une brève pression, les micro-aiguilles chargées de médicament peuvent être enfoncées dans la peau pour une libération du médicament.
PCT/SG2018/050282 2017-06-06 2018-06-06 Patch à micro-aiguilles chargé d'un agent de brunissement de graisse et son procédé de préparation WO2018226160A1 (fr)

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20200246321A1 (en) * 2017-09-13 2020-08-06 North Carolina State University Locally-induced adipose tissue browning by microneedle patch for obesity treatment
CN113332588A (zh) * 2021-05-26 2021-09-03 四川大学 用于口腔黏膜给药的尖端载药可溶性微针贴片及其制备方法
CN113332589A (zh) * 2021-05-26 2021-09-03 四川大学 用于口腔黏膜给药的装载双重药物的聚合物微针及其制备方法
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CN113133991B (zh) * 2020-01-19 2023-06-13 南京大学 秋水仙碱可溶性微针贴片及其制备方法
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US11793981B2 (en) 2020-05-21 2023-10-24 Equilibrate Therapeutics, LLC Microneedle device for control of thyroid hormone levels
CN113332588A (zh) * 2021-05-26 2021-09-03 四川大学 用于口腔黏膜给药的尖端载药可溶性微针贴片及其制备方法
CN113332589A (zh) * 2021-05-26 2021-09-03 四川大学 用于口腔黏膜给药的装载双重药物的聚合物微针及其制备方法

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