WO2020184131A1 - Systemically-acting external preparation containing sirolimus or derivative thereof - Google Patents

Systemically-acting external preparation containing sirolimus or derivative thereof Download PDF

Info

Publication number
WO2020184131A1
WO2020184131A1 PCT/JP2020/006722 JP2020006722W WO2020184131A1 WO 2020184131 A1 WO2020184131 A1 WO 2020184131A1 JP 2020006722 W JP2020006722 W JP 2020006722W WO 2020184131 A1 WO2020184131 A1 WO 2020184131A1
Authority
WO
WIPO (PCT)
Prior art keywords
sirolimus
external preparation
blood
preparation
weight
Prior art date
Application number
PCT/JP2020/006722
Other languages
French (fr)
Japanese (ja)
Inventor
眞理 金田
一朗 片山
Original Assignee
国立大学法人大阪大学
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 国立大学法人大阪大学 filed Critical 国立大学法人大阪大学
Publication of WO2020184131A1 publication Critical patent/WO2020184131A1/en

Links

Images

Classifications

    • 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
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/436—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a six-membered ring having oxygen as a ring hetero atom, e.g. rapamycin
    • 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
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7052—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides
    • A61K31/706—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom
    • 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
    • A61K47/44—Oils, fats or waxes according to two or more groups of A61K47/02-A61K47/42; Natural or modified natural oils, fats or waxes, e.g. castor oil, polyethoxylated castor oil, montan wax, lignite, shellac, rosin, beeswax or lanolin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders

Definitions

  • the present invention relates to an external preparation containing at least one selected from the group consisting of sirolimus and sirolimus derivatives.
  • Sirolimus and sirolimus derivatives have mTOR inhibitory activity, immunosuppressive activity, antiproliferative activity and the like.
  • Oral preparation containing sirolimus for lymphangioleiomyomatosis, topical sirolimus-containing preparation for skin lesions associated with tuberous sclerosis, and everolimus for tuberous sclerosis and some tumors Oral formulations are currently in use.
  • the present inventor has found that an external preparation containing sirolimus can treat skin lesions associated with tuberous sclerosis (Patent Document 1), and succeeded in developing and selling a safe external preparation with few side effects. ..
  • Sirolimus or sirolimus derivatives must be absorbed into the blood for treatment of lesions other than the skin and mucous membranes, but long-term administration by oral administration has caused side effects.
  • An object of the present invention is to provide an external preparation in which sirolimus or a sirolimus derivative is percutaneously or transmucosally absorbed into blood and exerts an action and effect on the whole body.
  • an external preparation containing at least one active ingredient selected from the group consisting of sirolimus and sirolimus derivatives and vegetable oil transfers the active ingredient into the blood. We found that we could do this, and completed the present invention.
  • the external preparation of the present invention can treat systemic lesions by rapidly transferring sirolimus or a sirolimus derivative into the blood.
  • the sirolimus or sirolimus derivative which is the active ingredient of the external preparation of the present invention, efficiently permeates the epidermis, passes through the dermis, and translocates into the blood.
  • the external preparation of the present invention can avoid the first pass effect and reduce the burden on the liver as compared with the oral preparation. It can also be administered to patients with dysphagia, patients who cannot take oral preparations temporarily or continuously due to impaired consciousness, convulsions, etc., and infants. It can be administered to elderly people with weak swallowing ability without causing aspiration.
  • the external preparation of the present invention can administer sirolimus or a sirolimus derivative into blood without pain like injection, and does not require administration technique.
  • the blood concentration rises within a few hours after administration and then falls, but in the case of an external preparation, a stable blood concentration can be maintained for a long time by the formulation design.
  • the external preparation of the present invention expands the application of sirolimus and sirolimus derivatives by developing a continuous preparation, optimizing the administration form according to the pathological condition, and the like.
  • FIG. 1 shows the sirolimus concentration in the dermis and blood at each time after a single transdermal administration of 0.4% sirolimus-containing liniment agent (SLN), which is an embodiment of the external preparation of the present invention, to mice. It is a graph.
  • SSN sirolimus-containing liniment agent
  • (A) indicates the concentration in the dermis (ng / mg) and
  • (b) indicates the concentration in the blood (ng / ml).
  • the insertion graph in (a) is an enlarged graph on the y-axis.
  • N 6, ** P ⁇ 0.01, * P ⁇ 0.05
  • FIG. 2 is a graph showing the sirolimus concentration in the dermis and blood at each time after a single transdermal administration of 0.8% sirolimus-containing gel (SG), which is a comparative preparation, to mice.
  • SG sirolimus-containing gel
  • (A) indicates the concentration in the dermis (ng / mg) and (b) indicates the concentration in the blood (ng / ml).
  • (N 6-9, ** P ⁇ 0.01)
  • FIG. 3 is a graph showing the sirolimus concentration in the dermis and blood at each time after oral administration of the comparative preparation sirolimus-containing oral preparation (Oral S) to mice.
  • (A) indicates the concentration in the dermis (ng / mg) and (b) indicates the concentration in the blood (ng / ml).
  • FIG. 4 is a graph showing the efficiency of delivery of sirolimus to the dermis and blood of a liniment agent (SLN), a gel agent (SG) and an oral preparation (Oral S) containing sirolimus.
  • SN liniment agent
  • SG gel agent
  • Oral S oral preparation
  • the external preparation according to the present invention contains at least one selected from the group consisting of sirolimus and sirolimus derivatives as an active ingredient.
  • the sirolimus derivative is not particularly limited, and examples thereof include everolimus, temsirolimus, lidaphorolimus, and zotarolimus. These have almost the same basic skeleton as the basic skeleton of sirolimus, and are known to have the same physiological activity as sirolimus. Therefore, these sirolimus derivatives can also be used as the active ingredient according to the embodiment of the present invention, like sirolimus.
  • Sirolimus and everolimus have already been used as therapeutic agents and have been confirmed to be clinically safe. Therefore, preferred active ingredients include sirolimus and everolimus. Sirolimus has already been used as an external preparation for the treatment of skin lesions associated with tuberous sclerosis, and a more preferable active ingredient is sirolimus.
  • the external preparation of the present invention is a preparation for transdermal or transmucosal administration, preferably a preparation for transdermal administration.
  • the dosage form of the external preparation of the present invention is not particularly limited. Examples thereof include liniments, ointments, patches, poultices, creams, gels, lotions, and suppositories. A liniment agent is preferred.
  • transdermal preparation which is a preferred embodiment of the external preparation of the present invention, can also be referred to as a transdermal preparation.
  • the transdermal preparation includes not only patches and poultices but also all external dosage forms such as liniments, ointments, creams, gels, lotions, and suppositories.
  • the amount of sirolimus or sirolimus derivative as an active ingredient contained in the external preparation of the present invention is not particularly limited.
  • the following concentrations are suitable from the viewpoint of rapidly migrating into the blood and obtaining a therapeutic effect.
  • the lower limit of the concentration of the active ingredient is 0.01% by weight or more, 0.02% by weight or more, 0.03% by weight or more, 0.04% by weight or more, 0.05% by weight based on the total weight of the external preparation. % Or more, 0.06% by weight or more, 0.07% by weight or more, 0.08% by weight or more, 0.09% by weight or more, 0.1% by weight or more.
  • the upper limit of the concentration of the active ingredient is 2.0% by weight or less, 1.5% by weight or less, 1.0% by weight or less, 0.9% by weight or less, 0.8% by weight based on the total weight of the external preparation. % Or less, 0.7% by weight or less, 0.6% by weight or less, 0.5% by weight or less, 0.4% by weight or less, 0.3% by weight or less, 0.25% by weight or less, 0.2% by weight It can be:
  • the concentration of the active ingredient is, for example, preferably 0.01 to 2.0% by weight, more preferably 0.03 to 1.0% by weight, and 0.04 to 0.8, based on the total weight of the external preparation. By weight% is more preferable, and 0.05 to 0.4% by weight is more preferable.
  • the daily dose of sirolimus or sirolimus derivative per unit surface area in a living body is not particularly limited.
  • the lower limit of the dosage per day 0.0001 mg / cm 2 or more, 0.0002 mg / cm 2 or more, 0.0005 mg / cm 2 or more, 0.001 mg / cm 2 or more, 0.002 mg / cm 2 As mentioned above, it can be 0.003 mg / cm 2 or more.
  • the upper limit of the dose is 2 mg / cm 2 or less, 1 mg / cm 2 or less, 0.5 mg / cm 2 or less, 0.1 mg / cm 2 or less, 0.05 mg / cm 2 or less, 0. 04mg / cm 2 or less, it may be 0.03 mg / cm 2 or less.
  • the dose is, for example, 0.0001 to 2 mg / cm 2 , preferably 0.0002 to 1 mg / cm 2 , more preferably 0.0005 to 0.5 mg / cm 2 , and more preferably 0.001 to 0. It is 1 mg / cm 2 .
  • the above-mentioned amount of sirolimus or sirolimus derivative may be administered once a day by application or the like, or may be administered in a plurality of times. In other words, it is preferable that the external preparation according to the present invention can realize the above-mentioned dose.
  • the external preparation of the present invention contains vegetable oil.
  • the vegetable oil is not particularly limited, but a preferable vegetable oil is edible oil.
  • the edible oil is not particularly limited, but preferred edible oils include sesame oil, olive oil, soybean oil, camellia oil and the like. From the viewpoint of skin permeability, sesame oil is most preferable. Further, edible oil heat-treated at 80 to 100 ° C. for 1 to 120 minutes is preferable, and sesame oil heat-treated at 80 to 100 ° C. for 1 to 120 minutes is more preferable.
  • the content of the vegetable oil contained in the external preparation of the present invention is, for example, preferably 10 to 99% by weight, more preferably 30 to 97% by weight, and even more preferably 50 to 95% by weight, based on the total weight of the external preparation. , 60-93% by weight is more preferable.
  • the external preparation of the present invention preferably contains alkanols C 2 ⁇ C 4.
  • Sirolimus may be admixed with a vegetable oil from dissolved in alkanol C 2 ⁇ C 4.
  • the alkanol C 2 ⁇ C 4 ethanol, isopropanol, n- propanol, butan-1-ol and butan-2-ol, and the like. Ethanol and isopropanol are preferred, and ethanol is most preferred. It may be a mixture of these C 2 to C 4 alkanols.
  • the amount of C 2 to C 4 alkanol contained in the external preparation of the present invention may be an amount capable of dissolving sirolimus or a sirolimus derivative.
  • 1 to 40% by weight is preferable, 2 to 30% by weight is more preferable, and 5 to 25% by weight is more preferable.
  • a liniment agent can be prepared by mixing a solution containing sirolimus or a sirolimus derivative with a vegetable oil.
  • An ointment can be prepared by mixing a solution containing sirolimus or a sirolimus derivative with a vegetable oil and then mixing with an ointment base.
  • Patches, poultices, lotions, gels and creams can be prepared according to well-known methods after mixing a solution containing sirolimus or a sirolimus derivative with vegetable oil.
  • Examples of the base and additives for the external preparation of the present invention include various components.
  • oily components oily components, aqueous components, surfactants, preservatives, pH adjusters, antioxidants and the like can be mentioned.
  • examples of the oily component include hydrocarbons, fatty acid esters, waxes, higher fatty acids, higher alcohols and the like.
  • it may contain a medicinal ingredient different from that of sirolimus or a sirolimus derivative.
  • compositions of the external preparation of the present invention include, for example, the following compositions based on the total weight of the external preparation.
  • Sirolimus or sirolimus derivative 0.01-2.0 wt%, vegetable oil 50-95 wt%, C 2 alkanol 5 ⁇ C 4 ⁇ 25 wt%
  • the external preparation of the present invention is an external preparation in which at least one selected from the group consisting of the active ingredients sirolimus and sirolimus derivatives is transferred into blood.
  • An external preparation that transfers into the blood is a preparation in which most of the active ingredients administered on the skin or mucous membrane pass through not only the skin or mucous membrane tissue but also into the blood in blood vessels. That is, the external preparation of the present invention is a preparation having excellent delivery efficiency of the active ingredient into blood.
  • the amount of the active ingredient present in the blood is 2 or more with respect to the amount of the active ingredient in the dermis under the applied region in 6 to 12 hours after administration. It is preferably 3 or more, more preferably 4 or more, and more preferably 5 or more, 6 or more, 7 or more, and 8 or more.
  • the external preparation of the present invention is an external preparation whose delivery efficiency of the active ingredient into blood is almost the same as that of an oral preparation.
  • AUC area under the blood drug concentration time curve
  • the AUC of the external preparation of the present invention is 0.1 or more, preferably 0.5 or more. , More preferably 0.8 or more, still more preferably 1 or more.
  • the external preparation of the present invention is administered to the affected area, for example, daily or once every 2 to 3 days.
  • Daily application is preferable, application 1 to 3 times a day is preferable, and application 2 to 3 times a day is more preferable.
  • an external preparation having a concentration of 0.05 to 0.2% may be applied 1 to 3 times a day.
  • 0.2 to 0.8% of an external preparation may be applied to the palm, footpad, etc. once or twice a day.
  • a liniment agent containing 0.4% by weight of sirolimus was prepared as follows.
  • Sirolimus powder was obtained from Fujian Kerui Pharmaceutical Co., Ltd.
  • Pretreated sesame oil was prepared by heating sesame oil at 95 ° C. for 5 minutes and then allowing it to cool to room temperature.
  • Sirolimus powder was added to ethanol to dissolve it and mixed with pretreated sesame oil to obtain a liniment agent.
  • the specific composition is shown below.
  • Example 2 A liniment agent containing 0.2% by weight of sirolimus was prepared in the same manner as in Example 1 except that the composition was changed to the following. The specific composition is shown below. Sirolimus 0.2g Ethanol 10g Sesame oil 89.8g (Total amount 100g)
  • Sirolimus-containing gel agent Sirolimus was added to ethanol to dissolve it, and then mixed with a pH-adjusted carboxyvinyl polymer to prepare a gel agent (SG) containing 0.8% of sirolimus.
  • the specific composition is shown below.
  • Sirolimus 0.8g Ethanol 48g Water for injection 49g Carbomer® 934P NF 1.6g Trishydroxymethylaminomethane 0.6g (Total amount 100g)
  • Oral preparation containing sirolimus A 1.25% sirolimus solution obtained by adding sirolimus to ethanol and dissolving it is mixed with a 9-fold amount of a physiological saline solution containing 0.1% Tween 20, and 1.25 mg of sirolimus in 1 ml. An oral preparation containing the above was obtained.
  • mice were injected intraperitoneally with an anesthetic (a combination of medetomidine 0.3 mg / kg, midazolam 4.0 mg / kg and butorphanol 5.0 mg / kg).
  • an E color (Natsume Seisakusho) was attached to the mouse to prevent grooming. The E-color was worn for up to 24 hours after administration of sirolimus.
  • Dermis and blood sampling Sampling was performed 1 hour, 3 hours, 6 hours, 12 hours, and 24 hours after the administration of sirolimus. Sampling was performed under sevoflurane anesthesia. The liniment or gel was removed by stripping with scotch tape several times. Blood was drawn from the inferior vena cava, internal jugular vein or cardiac puncture using an EDTA-coated syringe. Whole blood was inverted and mixed in a storage tube and stored at ⁇ 80 ° C. until measurement. After blood collection, the mice were sacrificed, the locally administered back skin was cut off, and the dermis was obtained with tweezers. Dermal tissue samples were snap frozen in liquid nitrogen and stored at -80 ° C until measurement.
  • sirolimus concentration in dermis and blood by LC-MS / MS was measured by the LC-MS / MS method using Waters ultra-high performance liquid chromatography (UPLC) and a tandem quadrupole mass spectrometer (TQD).
  • UPLC Waters ultra-high performance liquid chromatography
  • TQD tandem quadrupole mass spectrometer
  • the mass-to-charge ratio of silolimus used for quantification was m / z 931.7 (precursor ion) ⁇ 864.6 (product ion), and the mass-to-charge ratio of internal standard asomycin was m / z 809.7 (precursor ion) ⁇ 756.5 (product ion). Results were calculated using Waters' MassLynx 4.1 software. Tissue and blood sirolimus concentrations were calculated based on the tissue weight and blood volume used for quantification.
  • FIG. 1 (a) shows the sirolimus concentration in the dermis
  • FIG. 1 (b) shows the sirolimus concentration in the blood.
  • the concentration of sirolimus in the dermis was low and constant until after 12 hours, and became undetectable after 24 hours. Blood levels increased after 3 hours and increased after 6 and 12 hours.
  • FIG. 2 shows the sirolimus concentration in the dermis and the sirolimus concentration in the blood up to 24 hours after the single administration of the comparative preparation 0.8% sirolimus gel (SG) for external use.
  • FIG. 2A shows the sirolimus concentration in the dermis
  • FIG. 2B shows the sirolimus concentration in the blood.
  • FIG. 3 shows the sirolimus concentration in the dermis and the sirolimus concentration in the blood up to 24 hours after a single oral administration of the comparative oral preparation (Oral S).
  • FIG. 3 (a) shows the sirolimus concentration in the dermis
  • FIG. 3 (b) shows the sirolimus concentration in the blood. It hardly migrated into the dermis, and the concentration in blood decreased after 1 to 3 hours after the concentration increased.
  • Ds 50Cd / 10Sd
  • Ds Delivery efficiency of sirolimus to the dermis
  • Cd Median sirolimus concentration in the dermis (ng / mg)
  • Sd Dosing sirolimus amount ( ⁇ g)
  • Bs 2Cb / 10Sd
  • Bs Efficiency of delivery of sirolimus to blood
  • Cb Median blood sirolimus concentration (ng / mL)
  • Sd Dosing sirolimus amount ( ⁇ g) The results are shown in FIG.
  • the external preparation of the present invention is a preparation in which sirolimus efficiently permeates the epidermis and then transfers to the blood, not only in the dermis, and it has been shown that the efficiency of delivering sirolimus to the blood is equivalent to that of the oral preparation.
  • the external preparation of the present invention is a preparation that can be efficiently delivered into blood.
  • the external preparation of the present invention does not require administration technique, does not cause pain like injection, does not cause aspiration even in infants and elderly people with weak swallowing ability, and sirolimus or sirolimus derivative in blood. It can be administered to the patient and will greatly contribute to future medical care.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Public Health (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Epidemiology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Organic Chemistry (AREA)
  • Dermatology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Molecular Biology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicinal Preparation (AREA)

Abstract

[Problem] The present invention addresses the problem of providing an external preparation whereby sirolimus or a sirolimus derivative is absorbed transdermally or transmucosally into the blood and exerts an effect on the entire body. [Solution] Provided is an external preparation containing at least one selected from the group consisting of sirolimus and sirolimus derivatives, and vegetable oil. This external preparation has an active ingredient that passes through the skin or mucosa and migrates efficiently into the blood, and the rate of delivery of the active ingredient to the blood is substantially equivalent to that of an oral formulation.

Description

シロリムスまたはその誘導体を含有する全身作用型外用剤Systemic external preparation containing sirolimus or its derivative
本発明は、シロリムスおよびシロリムス誘導体からなる群より選択される少なくとも１つを含有する外用剤に関する。 The present invention relates to an external preparation containing at least one selected from the group consisting of sirolimus and sirolimus derivatives.
シロリムスおよびシロリムス誘導体はｍＴＯＲ阻害作用を有し、免疫抑制作用、抗増殖作用等を有する。リンパ脈管筋腫症を適応症とするシロリムス含有経口製剤、結節性硬化症に伴う皮膚病変を適応症とするシロリムス含有外用剤、および結節性硬化症といくつかの腫瘍を適応症とするエベロリムス含有経口製剤が現在使用されている。 Sirolimus and sirolimus derivatives have mTOR inhibitory activity, immunosuppressive activity, antiproliferative activity and the like. Oral preparation containing sirolimus for lymphangioleiomyomatosis, topical sirolimus-containing preparation for skin lesions associated with tuberous sclerosis, and everolimus for tuberous sclerosis and some tumors Oral formulations are currently in use.
本発明者は、シロリムスを含む外用剤により結節性硬化症に伴う皮膚病変の治療が可能であることを見出し（特許文献１）、副作用が少ない安全な外用剤を開発し販売することに成功した。 The present inventor has found that an external preparation containing sirolimus can treat skin lesions associated with tuberous sclerosis (Patent Document 1), and succeeded in developing and selling a safe external preparation with few side effects. ..
皮膚や粘膜以外に存在する病変部位へは、シロリムスまたはシロリムス誘導体を血液中に吸収させて治療しなければならないが、内服による長期間の投与では、副作用が問題になっていた。 Sirolimus or sirolimus derivatives must be absorbed into the blood for treatment of lesions other than the skin and mucous membranes, but long-term administration by oral administration has caused side effects.
国際公開２０１２／１０５５２１号公報International Publication No. 2012/105521
本発明は、シロリムスまたはシロリムス誘導体が経皮的または経粘膜的に血液中に吸収されて全身への作用効果を発揮する外用剤を提供することを課題とする。 An object of the present invention is to provide an external preparation in which sirolimus or a sirolimus derivative is percutaneously or transmucosally absorbed into blood and exerts an action and effect on the whole body.
本発明者は、前記課題に鑑み鋭意検討した結果、シロリムスおよびシロリムス誘導体からなる群より選択される少なくとも１つからなる有効成分と植物油を含有する外用剤が、有効成分を血液中に移行させることができることを見出し、本発明を完成するにいたった。 As a result of diligent studies in view of the above problems, the present inventor has found that an external preparation containing at least one active ingredient selected from the group consisting of sirolimus and sirolimus derivatives and vegetable oil transfers the active ingredient into the blood. We found that we could do this, and completed the present invention.
１．シロリムスおよびシロリムス誘導体からなる群より選択される少なくとも１つと植物油を含有する外用剤。
２．前記植物油が食用油である、前項１に記載の外用剤。
３．前記食用油が、ゴマ油、オリブ油、ダイズ油およびツバキ油からなる群から選択される少なくとも１つである、前項２に記載の外用剤。
４．前記食用油がゴマ油である、前項３に記載の外用剤。
５．さらに、Ｃ２～Ｃ４のアルカノールを含有する、前項１～４のいずれか１に記載の外用剤。
６．前記シロリムスおよびシロリムス誘導体からなる群より選択される少なくとも１つが血液中に移行する、前項１～５のいずれか１に記載の外用剤。
７．前記シロリムス誘導体が、エベロリムス、テムシロリムス、リダフォロリムスおよびゾタロリムスからなる群から選択される少なくとも１つである、前項１～６のいずれか１に記載の外用剤。 1. 1. An external preparation containing at least one selected from the group consisting of sirolimus and sirolimus derivatives and vegetable oil.
2. 2. The external preparation according to item 1 above, wherein the vegetable oil is an edible oil.
3. 3. The external preparation according to item 2 above, wherein the edible oil is at least one selected from the group consisting of sesame oil, olive oil, soybean oil and camellia oil.
4. The external preparation according to item 3 above, wherein the cooking oil is sesame oil.
5. Further, containing alkanols C 2 ~ C 4, external preparation according to any one of the preceding 1-4.
6. The external preparation according to any one of the above items 1 to 5, wherein at least one selected from the group consisting of sirolimus and sirolimus derivatives is transferred to blood.
7. The external preparation according to any one of items 1 to 6 above, wherein the sirolimus derivative is at least one selected from the group consisting of everolimus, temsirolimus, lidaphorolimus and zotarolimus.
本発明の外用剤は、シロリムスまたはシロリムス誘導体が速やかに血液中に移行することにより、全身の病変を治療することができる。本発明の外用剤の有効成分であるシロリムスまたはシロリムス誘導体は効率的に表皮に浸透し、真皮に留まらずに通過して血液中に移行する。また、本発明の外用剤は、経口製剤と比較すると、初回通過効果が回避でき肝臓の負担を少なくすることができる。また、嚥下障害がある患者、意識障害や痙攣等により一時的もしくは継続的に経口製剤を服用できない患者、および、乳児等に投与できる。嚥下能の弱った高齢者にも誤嚥を引き起こすことも無く投与できる。本発明の外用剤は、注射のような痛みを伴わずシロリムスまたはシロリムス誘導体を血中に投与でき、投与技術を必要としない。 The external preparation of the present invention can treat systemic lesions by rapidly transferring sirolimus or a sirolimus derivative into the blood. The sirolimus or sirolimus derivative, which is the active ingredient of the external preparation of the present invention, efficiently permeates the epidermis, passes through the dermis, and translocates into the blood. In addition, the external preparation of the present invention can avoid the first pass effect and reduce the burden on the liver as compared with the oral preparation. It can also be administered to patients with dysphagia, patients who cannot take oral preparations temporarily or continuously due to impaired consciousness, convulsions, etc., and infants. It can be administered to elderly people with weak swallowing ability without causing aspiration. The external preparation of the present invention can administer sirolimus or a sirolimus derivative into blood without pain like injection, and does not require administration technique.
また、経口投与では、投与後数時間で血中濃度が上昇しその後下降するが、外用剤では製剤設計により長時間安定した血中濃度を保つことができる。安定した血中濃度を保つことにより、より有効な効果を期待でき、副作用を軽減できると考えられる。本発明の外用剤は、持続製剤の開発、病態による投与形態の最適化等により、シロリムスおよびシロリムス誘導体の応用を広げるものである。 In addition, in oral administration, the blood concentration rises within a few hours after administration and then falls, but in the case of an external preparation, a stable blood concentration can be maintained for a long time by the formulation design. By maintaining a stable blood concentration, more effective effects can be expected and side effects can be reduced. The external preparation of the present invention expands the application of sirolimus and sirolimus derivatives by developing a continuous preparation, optimizing the administration form according to the pathological condition, and the like.
図１は、本発明の外用剤の一実施形態である０．４％シロリムス含有リニメント剤（ＳＬＮ）をマウスに単回経皮投与した後の各時間の真皮中および血液中のシロリムス濃度を示すグラフである。（ａ）は真皮中の濃度（ｎｇ／ｍｇ）（ｂ）は血液中の濃度（ｎｇ／ｍｌ）を示す。（ａ）の挿入グラフはｙ軸を拡大したグラフである。（Ｎ＝６、**Ｐ＜０．０１，*Ｐ＜０．０５）FIG. 1 shows the sirolimus concentration in the dermis and blood at each time after a single transdermal administration of 0.4% sirolimus-containing liniment agent (SLN), which is an embodiment of the external preparation of the present invention, to mice. It is a graph. (A) indicates the concentration in the dermis (ng / mg) and (b) indicates the concentration in the blood (ng / ml). The insertion graph in (a) is an enlarged graph on the y-axis. (N = 6, ** P <0.01, * P <0.05) 図２は、比較製剤である０．８％シロリムス含有ゲル剤（ＳＧ）をマウスに単回経皮投与した後の各時間の真皮中および血液中のシロリムス濃度を示すグラフである。（ａ）は真皮中の濃度（ｎｇ／ｍｇ）（ｂ）は血液中の濃度（ｎｇ／ｍｌ）を示す。（Ｎ＝６－９、**Ｐ＜０．０１）FIG. 2 is a graph showing the sirolimus concentration in the dermis and blood at each time after a single transdermal administration of 0.8% sirolimus-containing gel (SG), which is a comparative preparation, to mice. (A) indicates the concentration in the dermis (ng / mg) and (b) indicates the concentration in the blood (ng / ml). (N = 6-9, ** P <0.01) 図３は、比較製剤であるシロリムス含有経口製剤（Ｏｒａｌ　Ｓ）をマウスに経口投与した後の各時間の真皮中および血液中のシロリムス濃度を示すグラフである。（ａ）は真皮中の濃度（ｎｇ／ｍｇ）（ｂ）は血液中の濃度（ｎｇ／ｍｌ）を示す。（ａ）の挿入グラフはｙ軸を拡大したグラフである。（Ｎ＝６－７、**Ｐ＜０．０１）FIG. 3 is a graph showing the sirolimus concentration in the dermis and blood at each time after oral administration of the comparative preparation sirolimus-containing oral preparation (Oral S) to mice. (A) indicates the concentration in the dermis (ng / mg) and (b) indicates the concentration in the blood (ng / ml). The insertion graph in (a) is an enlarged graph on the y-axis. (N = 6-7, ** P <0.01) 図４は、シロリムスを含有するリニメント剤（ＳＬＮ）、ゲル剤（ＳＧ）および経口製剤（Ｏｒａｌ　Ｓ）の真皮および血液へのシロリムス送達効率を示すグラフである。各シロリムス製剤の単回投与後の各時間に、シロリムス投与量の何％が真皮中および血液中に存在するか（％用量）を示す。FIG. 4 is a graph showing the efficiency of delivery of sirolimus to the dermis and blood of a liniment agent (SLN), a gel agent (SG) and an oral preparation (Oral S) containing sirolimus. At each time after a single dose of each sirolimus formulation, what percentage of the sirolimus dose is present in the dermis and blood (% dose) is shown.
本発明は以下に説明した実施形態に限定されるものではなく、特許請求の範囲に示した範囲において種々の変更が可能である。異なる実施形態や実施例に、それぞれ開示された技術的手段を適宜組み合わせて得られる実施形態や実施例についても、本発明の技術的範囲に含まれる。 The present invention is not limited to the embodiments described below, and various modifications can be made within the scope of the claims. The technical scope of the present invention also includes embodiments and examples obtained by appropriately combining the disclosed technical means with different embodiments and examples.
本発明に係る外用剤は、シロリムスおよびシロリムス誘導体からなる群より選択される少なくとも１つを有効成分として含有する。 The external preparation according to the present invention contains at least one selected from the group consisting of sirolimus and sirolimus derivatives as an active ingredient.
前記シロリムス誘導体は、特に限定されないが、例えばエベロリムス、テムシロリムス、リダフォロリムスおよびゾタロリムスを挙げることができる。これらは、シロリムスの基本骨格とほぼ同じ基本骨格を有しており、シロリムスと同等の生理活性を有することが知られている。したがって、これらのシロリムス誘導体も、シロリムスと同様に、本発明の一実施形態に係る有効成分として用いることができる。 The sirolimus derivative is not particularly limited, and examples thereof include everolimus, temsirolimus, lidaphorolimus, and zotarolimus. These have almost the same basic skeleton as the basic skeleton of sirolimus, and are known to have the same physiological activity as sirolimus. Therefore, these sirolimus derivatives can also be used as the active ingredient according to the embodiment of the present invention, like sirolimus.
シロリムスおよびエベロリムスは、治療薬としてすでに用いられており、臨床における安全性が確認されている。よって、好ましい有効成分としては、シロリムスおよびエベロリムスが挙げられる。シロリムスは、結節性硬化症に伴う皮膚病変の治療のための外用薬としてもすでに用いられており、より好ましい有効成分はシロリムスである。 Sirolimus and everolimus have already been used as therapeutic agents and have been confirmed to be clinically safe. Therefore, preferred active ingredients include sirolimus and everolimus. Sirolimus has already been used as an external preparation for the treatment of skin lesions associated with tuberous sclerosis, and a more preferable active ingredient is sirolimus.
本発明の外用剤は、経皮または経粘膜投与用製剤であり、好ましくは経皮投与用製剤である。本発明の外用剤の剤形は特に限定されない。リニメント剤、軟膏剤、貼付剤、パップ剤、クリーム剤、ゲル剤、ローション剤、および、坐剤等が挙げられる。好ましくは、リニメント剤である。 The external preparation of the present invention is a preparation for transdermal or transmucosal administration, preferably a preparation for transdermal administration. The dosage form of the external preparation of the present invention is not particularly limited. Examples thereof include liniments, ointments, patches, poultices, creams, gels, lotions, and suppositories. A liniment agent is preferred.
本発明の外用剤の好ましい態様である経皮投与用製剤は経皮吸収型製剤ともいうことができる。本明細書において経皮吸収型製剤は、貼付剤、パップ剤だけでなくリニメント剤、軟膏剤、クリーム剤、ゲル剤、ローション剤、および、坐剤等の外用のすべての剤形を含む。 The preparation for transdermal administration, which is a preferred embodiment of the external preparation of the present invention, can also be referred to as a transdermal preparation. In the present specification, the transdermal preparation includes not only patches and poultices but also all external dosage forms such as liniments, ointments, creams, gels, lotions, and suppositories.
本発明の外用剤に含有される有効成分であるシロリムスまたはシロリムス誘導体の量は、特に限定されない。血液中へ速やかに移行し、治療効果を得るという観点から、以下の濃度が好適である。 The amount of sirolimus or sirolimus derivative as an active ingredient contained in the external preparation of the present invention is not particularly limited. The following concentrations are suitable from the viewpoint of rapidly migrating into the blood and obtaining a therapeutic effect.
前記有効成分の濃度の下限は、外用剤の総重量を基準として、０．０１重量％以上、０．０２重量％以上、０．０３重量％以上、０．０４重量％以上、０．０５重量％以上、０．０６重量％以上、０．０７重量％以上、０．０８重量％以上、０．０９重量％以上、０．１重量％以上でありうる。  The lower limit of the concentration of the active ingredient is 0.01% by weight or more, 0.02% by weight or more, 0.03% by weight or more, 0.04% by weight or more, 0.05% by weight based on the total weight of the external preparation. % Or more, 0.06% by weight or more, 0.07% by weight or more, 0.08% by weight or more, 0.09% by weight or more, 0.1% by weight or more.
前記有効成分の濃度の上限は、外用剤の総重量を基準として、２．０重量％以下、１．５重量％以下、１．０重量％以下、０．９重量％以下、０．８重量％以下、０．７重量％以下、０．６重量％以下、０．５重量％以下、０．４重量％以下、０．３重量％以下、０．２５重量％以下、０．２重量％以下でありうる。  The upper limit of the concentration of the active ingredient is 2.0% by weight or less, 1.5% by weight or less, 1.0% by weight or less, 0.9% by weight or less, 0.8% by weight based on the total weight of the external preparation. % Or less, 0.7% by weight or less, 0.6% by weight or less, 0.5% by weight or less, 0.4% by weight or less, 0.3% by weight or less, 0.25% by weight or less, 0.2% by weight It can be:
前記有効成分の濃度は、例えば、外用剤の総重量を基準として、０．０１～２．０重量％が好ましく、０．０３～１．０重量％がより好ましく、０．０４～０．８重量％がより好ましく０．０５～０．４重量％がより好ましい。 The concentration of the active ingredient is, for example, preferably 0.01 to 2.0% by weight, more preferably 0.03 to 1.0% by weight, and 0.04 to 0.8, based on the total weight of the external preparation. By weight% is more preferable, and 0.05 to 0.4% by weight is more preferable.
生体における単位表面積あたりの、シロリムスまたはシロリムス誘導体の１日あたりの投与量は特に限定されない。  The daily dose of sirolimus or sirolimus derivative per unit surface area in a living body is not particularly limited.
例えば、前記投与量の下限は、１日あたり０．０００１ｍｇ／ｃｍ２以上、０．０００２ｍｇ／ｃｍ２以上、０．０００５ｍｇ／ｃｍ２以上、０．００１ｍｇ／ｃｍ２以上、０．００２ｍｇ／ｃｍ２以上、０．００３ｍｇ／ｃｍ２以上でありうる。  For example, the lower limit of the dosage per day 0.0001 mg / cm 2 or more, 0.0002 mg / cm 2 or more, 0.0005 mg / cm 2 or more, 0.001 mg / cm 2 or more, 0.002 mg / cm 2 As mentioned above, it can be 0.003 mg / cm 2 or more.
例えば、前記投与量の上限は、１日あたり２ｍｇ／ｃｍ２以下、１ｍｇ／ｃｍ２以下、０．５ｍｇ／ｃｍ２以下、０．１ｍｇ／ｃｍ２以下、０．０５ｍｇ／ｃｍ２以下、０．０４ｍｇ／ｃｍ２以下、０．０３ｍｇ／ｃｍ２以下でありうる。  For example, the upper limit of the dose is 2 mg / cm 2 or less, 1 mg / cm 2 or less, 0.5 mg / cm 2 or less, 0.1 mg / cm 2 or less, 0.05 mg / cm 2 or less, 0. 04mg / cm 2 or less, it may be 0.03 mg / cm 2 or less.
前記投与量は、例えば、０．０００１～２ｍｇ／ｃｍ２、好ましくは０．０００２～１ｍｇ／ｃｍ２、より好ましくは０．０００５～０．５ｍｇ／ｃｍ２、より好ましくは０．００１～０．１ｍｇ／ｃｍ２、である。 The dose is, for example, 0.0001 to 2 mg / cm 2 , preferably 0.0002 to 1 mg / cm 2 , more preferably 0.0005 to 0.5 mg / cm 2 , and more preferably 0.001 to 0. It is 1 mg / cm 2 .
上述した量のシロリムスまたはシロリムス誘導体を、塗布などにより１日に１回投与、または複数回に分けて投与すればよい。換言すれば、本発明に係る外用剤は、上述した投与量を実現できるものであることが好ましい。 The above-mentioned amount of sirolimus or sirolimus derivative may be administered once a day by application or the like, or may be administered in a plurality of times. In other words, it is preferable that the external preparation according to the present invention can realize the above-mentioned dose.
本発明の外用剤は、植物油を含有する。植物油は特に限定されるものではないが、好ましい植物油としては食用油が挙げられる。食用油は特に限定されるものではないが、好ましい食用油としては、ゴマ油、オリブ油、ダイズ油およびツバキ油等が挙げられる。皮膚透過性の観点から最も好ましくは、ゴマ油である。また、８０～１００℃で１～１２０分熱処理された食用油が好ましく、８０～１００℃で１～１２０分熱処理されたゴマ油がより好ましい。 The external preparation of the present invention contains vegetable oil. The vegetable oil is not particularly limited, but a preferable vegetable oil is edible oil. The edible oil is not particularly limited, but preferred edible oils include sesame oil, olive oil, soybean oil, camellia oil and the like. From the viewpoint of skin permeability, sesame oil is most preferable. Further, edible oil heat-treated at 80 to 100 ° C. for 1 to 120 minutes is preferable, and sesame oil heat-treated at 80 to 100 ° C. for 1 to 120 minutes is more preferable.
本発明の外用剤に含まれる植物油の含有量は、外用剤の総重量を基準として、例えば、１０～９９重量％が好ましく、３０～９７重量％がより好ましく、５０～９５重量％がより好ましく、６０～９３重量％がさらに好ましい。 The content of the vegetable oil contained in the external preparation of the present invention is, for example, preferably 10 to 99% by weight, more preferably 30 to 97% by weight, and even more preferably 50 to 95% by weight, based on the total weight of the external preparation. , 60-93% by weight is more preferable.
本発明の外用剤は、好適には、Ｃ２～Ｃ４のアルカノールを含有する。シロリムスをＣ２～Ｃ４のアルカノールに溶解させてから植物油と混合することができる。Ｃ２～Ｃ４のアルカノールとしては、エタノール、イソプロパノール、ｎ‐プロパノール、ブタン‐１‐オールおよびブタン‐２‐オール等が挙げられる。好ましくは、エタノールおよびイソプロパノールであり、最も好ましくはエタノールである。これらＣ２～Ｃ４のアルカノールの混合物であってもよい。 The external preparation of the present invention preferably contains alkanols C 2 ~ C 4. Sirolimus may be admixed with a vegetable oil from dissolved in alkanol C 2 ~ C 4. The alkanol C 2 ~ C 4, ethanol, isopropanol, n- propanol, butan-1-ol and butan-2-ol, and the like. Ethanol and isopropanol are preferred, and ethanol is most preferred. It may be a mixture of these C 2 to C 4 alkanols.
本発明の外用剤に含まれるＣ２～Ｃ４のアルカノールの量は、シロリムスまたはシロリムス誘導体を溶解できる量であればよい。例えば、外用剤の総重量を基準として、１～４０重量％が好ましく、２～３０重量％がより好ましく、５～２５重量％がより好ましい。 The amount of C 2 to C 4 alkanol contained in the external preparation of the present invention may be an amount capable of dissolving sirolimus or a sirolimus derivative. For example, based on the total weight of the external preparation, 1 to 40% by weight is preferable, 2 to 30% by weight is more preferable, and 5 to 25% by weight is more preferable.
本発明の外用剤の調製方法として、以下のものが挙げられる。（ｉ）シロリムスまたはシロリムス誘導体を含有している溶液を植物油と混合することによりリニメント剤を調製しうる。（ｉｉ）シロリムスまたはシロリムス誘導体を含有している溶液を植物油と混合した後、軟膏基剤と混合することにより軟膏剤を調製しうる。（ｉｉｉ）貼付剤、パップ剤、ローション剤、ゲル剤およびクリーム剤は、シロリムスまたはシロリムス誘導体を含有している溶液を植物油と混合した後に周知の方法に従って調製しうる。  Examples of the method for preparing the external preparation of the present invention include the following. (I) A liniment agent can be prepared by mixing a solution containing sirolimus or a sirolimus derivative with a vegetable oil. (Ii) An ointment can be prepared by mixing a solution containing sirolimus or a sirolimus derivative with a vegetable oil and then mixing with an ointment base. (Iii) Patches, poultices, lotions, gels and creams can be prepared according to well-known methods after mixing a solution containing sirolimus or a sirolimus derivative with vegetable oil.
本発明の外用剤の基剤および添加剤としては、種々の成分が挙げられる。たとえば、油性成分、水性成分、界面活性剤、保存剤、ｐＨ調整剤、抗酸化剤等が挙げられる。油性成分としては、炭化水素類、脂肪酸エステル類、ロウ類、高級脂肪酸および高級アルコール等が挙げられる。また、シロリムスまたはシロリムス誘導体とは異なる薬効成分を含有してもよい。 Examples of the base and additives for the external preparation of the present invention include various components. For example, oily components, aqueous components, surfactants, preservatives, pH adjusters, antioxidants and the like can be mentioned. Examples of the oily component include hydrocarbons, fatty acid esters, waxes, higher fatty acids, higher alcohols and the like. In addition, it may contain a medicinal ingredient different from that of sirolimus or a sirolimus derivative.
本発明の外用剤の好ましい組成は、外用剤の総重量を基準として例えば以下のような組成が挙げられる。シロリムスまたはシロリムス誘導体０．０１～２．０重量％、植物油５０～９５重量％、Ｃ２～Ｃ４のアルカノール５～２５重量％ Preferred compositions of the external preparation of the present invention include, for example, the following compositions based on the total weight of the external preparation. Sirolimus or sirolimus derivative 0.01-2.0 wt%, vegetable oil 50-95 wt%, C 2 alkanol 5 ~ C 4 ~ 25 wt%
本発明の外用剤は、有効成分である前記シロリムスおよびシロリムス誘導体からなる群より選択される少なくとも１つが血液中に移行する外用剤である。血液中に移行する外用剤とは、皮膚や粘膜上に投与された有効成分の多くが、皮膚や粘膜組織に留まらず通過して血管内の血液中に移行する製剤である。すなわち、本発明の外用剤は、血液中への有効成分の送達効率に優れた製剤である。具体的には、本発明の外用剤は、投与後6～１２時間において、塗布した領域下の真皮中の有効成分の量に対する血中に存在する有効成分の量が２以上である。好ましくは３以上であり、さらに好ましくは４以上であり、より好ましくは５以上、６以上、７以上、８以上である。 The external preparation of the present invention is an external preparation in which at least one selected from the group consisting of the active ingredients sirolimus and sirolimus derivatives is transferred into blood. An external preparation that transfers into the blood is a preparation in which most of the active ingredients administered on the skin or mucous membrane pass through not only the skin or mucous membrane tissue but also into the blood in blood vessels. That is, the external preparation of the present invention is a preparation having excellent delivery efficiency of the active ingredient into blood. Specifically, in the external preparation of the present invention, the amount of the active ingredient present in the blood is 2 or more with respect to the amount of the active ingredient in the dermis under the applied region in 6 to 12 hours after administration. It is preferably 3 or more, more preferably 4 or more, and more preferably 5 or more, 6 or more, 7 or more, and 8 or more.
本発明の外用剤は、血液中への有効成分の送達効率が経口製剤とほぼ同等である外用剤である。ＡＵＣ（血中薬物濃度時間曲線下面積）で比較した場合、経口投与製剤単回投与のＡＵＣを１とすると、本発明の外用剤のＡＵＣは０．１以上であり、好ましくは０．５以上、さらに好ましくは０．８以上、さらに好ましくは、１以上である。 The external preparation of the present invention is an external preparation whose delivery efficiency of the active ingredient into blood is almost the same as that of an oral preparation. When compared by AUC (area under the blood drug concentration time curve), assuming that the AUC of a single dose of the oral preparation is 1, the AUC of the external preparation of the present invention is 0.1 or more, preferably 0.5 or more. , More preferably 0.8 or more, still more preferably 1 or more.
本発明の外用剤の投与は、例えば、毎日、または、２～３日に１回患部に塗布する。毎日の塗布が好ましく、１日に１～３回の塗布が好ましく、１日に２～３回の塗布がさらに好ましい。 The external preparation of the present invention is administered to the affected area, for example, daily or once every 2 to 3 days. Daily application is preferable, application 1 to 3 times a day is preferable, and application 2 to 3 times a day is more preferable.
また、疾患、症状、年齢、投与部位、皮膚の厚さ等により、外用剤中の有効成分の濃度および投与回数を調整することが好ましい。例えば小さい子供の薄い皮膚や腋下等には、０．０５～０．２％の濃度の外用剤を１日に１～３回塗布する態様が挙げられる。また、手掌、足蹠等には、０．２～０．８％の外用剤を１日に１～２回塗布する態様が挙げられる。 In addition, it is preferable to adjust the concentration of the active ingredient in the external preparation and the number of administrations according to the disease, symptoms, age, administration site, skin thickness and the like. For example, on the thin skin and armpits of small children, an external preparation having a concentration of 0.05 to 0.2% may be applied 1 to 3 times a day. In addition, 0.2 to 0.8% of an external preparation may be applied to the palm, footpad, etc. once or twice a day.
以下に、実施例を挙げて本発明をさらに具体的に説明するが、本発明はこれらに限定されるものではない。 Hereinafter, the present invention will be described in more detail with reference to examples, but the present invention is not limited thereto.
[実施例１]
シロリムスを０．４重量％含有するリニメント剤を次のように調製した。シロリムス粉末は、Fujian Kerui Pharmaceutical Co., Ltdより入手した。ゴマ油を９５℃で５分間加熱後放置して室温まで冷却し前処理ゴマ油を準備した。シロリムス粉末をエタノールに添加して溶解し、前処理ゴマ油と混合しリニメント剤を得た。具体的な組成を下記に示す。
　シロリムス　０．４ｇ
　エタノール　１０ｇ
　ゴマ油　８９．６ｇ
　（全量１００ｇ） [Example 1]
A liniment agent containing 0.4% by weight of sirolimus was prepared as follows. Sirolimus powder was obtained from Fujian Kerui Pharmaceutical Co., Ltd. Pretreated sesame oil was prepared by heating sesame oil at 95 ° C. for 5 minutes and then allowing it to cool to room temperature. Sirolimus powder was added to ethanol to dissolve it and mixed with pretreated sesame oil to obtain a liniment agent. The specific composition is shown below.
Sirolimus 0.4g
Ethanol 10g
Sesame oil 89.6g
(Total amount 100g)
[実施例２]
組成を下記に変更した以外は実施例１と同様にして、シロリムスを０．２重量％含有するリニメント剤を調製した。具体的な組成を下記に示す。
　シロリムス　０．２ｇ
　エタノール　１０ｇ
　ゴマ油　８９．８ｇ
　（全量１００ｇ） [Example 2]
A liniment agent containing 0.2% by weight of sirolimus was prepared in the same manner as in Example 1 except that the composition was changed to the following. The specific composition is shown below.
Sirolimus 0.2g
Ethanol 10g
Sesame oil 89.8g
(Total amount 100g)
[試験例] [Test example]
１．比較製剤の調製 1. 1. Preparation of comparative formulation
１－１．シロリムス含有ゲル剤
シロリムスをエタノールに添加して溶解した後、ｐＨを調整したカルボキシビニルポリマーと混合し、シロリムスを０．８％含有するゲル剤（ＳＧ）を調製した。具体的組成を以下に示す。
　シロリムス ０．８ｇ
　エタノール ４８ｇ
　注射用水 ４９ｇ
　カーボポール（登録商標）９３４Ｐ ＮＦ １．６ｇ
　トリスヒドロキシメチルアミノメタン ０．６ｇ
　（全量１００ｇ） 1-1. Sirolimus-containing gel agent Sirolimus was added to ethanol to dissolve it, and then mixed with a pH-adjusted carboxyvinyl polymer to prepare a gel agent (SG) containing 0.8% of sirolimus. The specific composition is shown below.
Sirolimus 0.8g
Ethanol 48g
Water for injection 49g
Carbomer® 934P NF 1.6g
Trishydroxymethylaminomethane 0.6g
(Total amount 100g)
１－２．シロリムス含有経口製剤
シロリムスをエタノールに添加して溶解して得た１．２５％シロリムス溶液を９倍量の０．１％ Ｔｗｅｅｎ ２０を含有する生理食塩液と混合し、１ｍｌに１．２５ｍｇのシロリムスを含有する経口製剤を得た。 1-2. Oral preparation containing sirolimus A 1.25% sirolimus solution obtained by adding sirolimus to ethanol and dissolving it is mixed with a 9-fold amount of a physiological saline solution containing 0.1% Tween 20, and 1.25 mg of sirolimus in 1 ml. An oral preparation containing the above was obtained.
２．実験動物
７週齢の雄のマウス（Ｈｏｓ：ＨＲ－１　ヘアレスマウス）を星野試験動物飼育所より購入し、５日間飼育した後試験を開始した。 2. 2. Experimental animals A 7-week-old male mouse (Hos: HR-1 hairless mouse) was purchased from the Hoshino Test Animal Breeding Center and bred for 5 days before starting the test.
３．シロリムス製剤の投与 3. 3. Administration of sirolimus preparation
３－１．経口投与
経口ゾンデ（０．９×５０ｍｍ、夏目製作所）を用いて上記シロリムス経口製剤０．２ｍｌを上記マウスに１回投与した（Ｎ＝６または７）。 3-1. Oral administration 0.2 ml of the above sirolimus oral preparation was administered once to the above mice using an oral sonde (0.9 × 50 mm, Natsume Seisakusho) (N = 6 or 7).
３－２．局所投与
上記マウスに、麻酔薬（メデトミジン０．３ｍｇ／ｋｇ、ミダゾラム４．０ｍｇ／ｋｇおよびブトルファノール５．０ｍｇ／ｋｇの組合せ）を、腹膜内に注射した。また、グルーミングを防止するためのＥカラー（夏目製作所）をマウスに装着した。Ｅカラーは最長でシロリムス投与後２４時間まで装着させた。上記実施例１の０．４％シロリムス含有リニメント剤１２５ｍｇを、マウス背部皮膚（２．５×３．５ｃｍ）に１回塗布した（Ｎ＝６）。また、上記比較製剤の０．８％シロリムス含有ゲル剤（ＳＧ）１２５ｍｇを、マウス背部皮膚（２．５×３．５ｃｍ）に１回塗布した（Ｎ＝６－９）。 3-2. Topical administration The above mice were injected intraperitoneally with an anesthetic (a combination of medetomidine 0.3 mg / kg, midazolam 4.0 mg / kg and butorphanol 5.0 mg / kg). In addition, an E color (Natsume Seisakusho) was attached to the mouse to prevent grooming. The E-color was worn for up to 24 hours after administration of sirolimus. 125 mg of the 0.4% sirolimus-containing liniment agent of Example 1 was applied once to the back skin (2.5 × 3.5 cm) of a mouse (N = 6). In addition, 125 mg of a 0.8% sirolimus-containing gel (SG) of the above comparative preparation was applied once to the back skin (2.5 × 3.5 cm) of a mouse (N = 6-9).
４．真皮と血液のサンプリング
シロリムス投与から、１時間後、３時間後、６時間後、１２時間後、２４時間後にサンプリングした。サンプリングはセボフルラン麻酔下でおこなった。リニメント剤またはゲル剤を数回スコッチテープでストリッピングすることによって取り除いた。採血は、ＥＤＴＡ被覆シリンジを使用して下大静脈、内頚静脈または心臓穿刺からおこなった。保存チューブ内で全血を転倒混和し、測定まで－８０℃で保存した。採血後、マウスを屠殺し、局所投薬した背部皮膚を切り取りピンセットで真皮を取得した。真皮の組織サンプルは、液体窒素で急速冷凍し測定まで－８０℃で保存した。 4. Dermis and blood sampling Sampling was performed 1 hour, 3 hours, 6 hours, 12 hours, and 24 hours after the administration of sirolimus. Sampling was performed under sevoflurane anesthesia. The liniment or gel was removed by stripping with scotch tape several times. Blood was drawn from the inferior vena cava, internal jugular vein or cardiac puncture using an EDTA-coated syringe. Whole blood was inverted and mixed in a storage tube and stored at −80 ° C. until measurement. After blood collection, the mice were sacrificed, the locally administered back skin was cut off, and the dermis was obtained with tweezers. Dermal tissue samples were snap frozen in liquid nitrogen and stored at -80 ° C until measurement.
５．ＬＣ－ＭＳ／ＭＳのためのシロリムス抽出物の調製 5. Preparation of sirolimus extract for LC-MS / MS
５－１．真皮サンプルからの調製
上記サンプリングで得た冷凍組織（およそ２０～３０ｍｇ）の組織重量を微量電子天秤で測定後、マルチビーズショッカー（登録商標）（安井器械）を用いて２８００ｒｐｍ、１５秒の条件下で破砕した。そして、以下の方法でシロリムス抽出をおこなった。冷却メタノール７００μｌを破砕組織に加え、その混合物をマイクロチューブへ移した。メタノール１００μｌを加え、シロリムスを抽出するために、１分間ボルテックスにかけた。４℃、１３，０００ｇ、５分で遠心分離した上清をサンプルとした。１ｎｇのアスコマイシン内部標準を加えた後に、ＬＣ－ＭＳ／ＭＳ分析のために用いた。未処理のＨＲマウス背部皮膚を同様に破砕調整したメタノール混合物にシロリムス標品１００μｌを加えた抽出サンプルを較正標準に用いた。 5-1. Preparation from dermis sample After measuring the tissue weight of the frozen tissue (approximately 20 to 30 mg) obtained by the above sampling with a microelectronic balance, a multi-bead shocker (registered trademark) (Yasui Kikai) was used at 2800 rpm for 15 seconds. Crushed in. Then, sirolimus was extracted by the following method. 700 μl of chilled methanol was added to the disrupted structure and the mixture was transferred to a microtube. 100 μl of methanol was added and vortexed for 1 minute to extract sirolimus. The supernatant obtained by centrifuging at 4 ° C., 13,000 g and 5 minutes was used as a sample. It was used for LC-MS / MS analysis after adding 1 ng of Ascomycin internal standard. An extracted sample was used as a calibration standard in which 100 μl of sirolimus preparation was added to a methanol mixture obtained by similarly crushing and adjusting the back skin of untreated HR mice.
５－２．血液サンプルからの調製
合計１．５ｎｇのアスコマイシン内部標準を、収集された全血１００μｌに加え、そして、０．２Ｍの硫酸亜鉛含有７０％メタノール３００μｌを加えた。得られた溶液を５分間ボルテックスにかけた後、６０００ｒｐｍで５分間遠心分離した。得られた上清をろ過し（ポアサイズ０．４５μｍ）、ＬＣ－ＭＳ／ＭＳ分析のために用いた。未処置ＨＲマウスからの血液が、較正標準のために使われた。 5-2. Preparation from Blood Samples A total of 1.5 ng of Ascomycin internal standard was added to 100 μl of collected whole blood and 300 μl of 0.2 M zinc sulphate-containing 70% methanol was added. The resulting solution was vortexed for 5 minutes and then centrifuged at 6000 rpm for 5 minutes. The resulting supernatant was filtered (pore size 0.45 μm) and used for LC-MS / MS analysis. Blood from untreated HR mice was used for calibration standards.
６．ＬＣ－ＭＳ／ＭＳによる真皮中および血液中のシロリムス濃度の評価
Waters社製超高速液体クロマトグラフィー (UPLC) とタンデム四重極型質量分析装置 (TQD) によるLC-MS/MS法によりシロリムス濃度測定を行なった。超高速液体クロマトグラフィーによるシロリムスサンプルの分離にはオクタデシルカラム、移動相Aに0.1%ギ酸、20 mM 酢酸アンモニウムを添加した超純水、移動相Bに0.1%ギ酸、20 mM酢酸アンモニウムを添加したメタノールを用いた。シロリムスサンプルのオクタデシルカラムからの分離・溶出は流速0.25 mL/分、移動相A : 移動相B を以下の濃度勾配で変動させることにより行い、総測定時間は10分間であった。移動相A : 移動相B = 50% : 50%からスタートし最初の２分間で移動相A : 移動相B = 0% : 100%に達するように勾配をかけ、移動相B: 100%の状態を５分継続、0.1分の間に移動相A : 移動相B = 50% : 50%に戻しそのまま残り2.9分間平衡状態にした。分離したシロリムスサンプルはエレクトロスプレーイオン化法(ESI positive mode)により定量した。定量に用いたシロリムスの質量電荷比はm/z 931.7 (プリカーサーイオン)< 864.6 (プロダクトイオン)、内部標準アスコマイシンの質量電荷比はm/z 809.7 (プリカーサーイオン)<756.5 (プロダクトイオン)とし、結果はWaters社のMassLynx4.1ソフトによって計算した。定量に用いた組織重量・血液量をもとに組織および血中シロリムス濃度を算出した。 6. Evaluation of sirolimus concentration in dermis and blood by LC-MS / MS
The sirolimus concentration was measured by the LC-MS / MS method using Waters ultra-high performance liquid chromatography (UPLC) and a tandem quadrupole mass spectrometer (TQD). For separation of silolimus samples by ultra-high performance liquid chromatography, octadecyl column, ultrapure water with 0.1% formic acid and 20 mM ammonium acetate added to mobile phase A, and methanol with 0.1% formic acid and 20 mM ammonium acetate added to mobile phase B. Was used. Separation and elution of the sirolimus sample from the octadecyl column was performed by varying the flow rate of 0.25 mL / min and mobile phase A: mobile phase B with the following concentration gradient, and the total measurement time was 10 minutes. Mobile phase A: Mobile phase B = 50%: Start from 50% and in the first 2 minutes Mobile phase A: Mobile phase B = 0%: Gradient to reach 100%, Mobile phase B: 100% state Was continued for 5 minutes, and within 0.1 minutes, mobile phase A: mobile phase B = 50%: 50% was returned to the equilibrium state for the remaining 2.9 minutes. The separated sirolimus samples were quantified by electrospray ionization (ESI positive mode). The mass-to-charge ratio of silolimus used for quantification was m / z 931.7 (precursor ion) <864.6 (product ion), and the mass-to-charge ratio of internal standard asomycin was m / z 809.7 (precursor ion) <756.5 (product ion). Results were calculated using Waters' MassLynx 4.1 software. Tissue and blood sirolimus concentrations were calculated based on the tissue weight and blood volume used for quantification.
７．結果
７－１．真皮中および血液中のシロリムスの濃度
実施例１の０．４％シロリムス含有リニメント剤（ＳＬＮ）を外用で単回投与した時の２４時間後までの真皮中のシロリムス濃度と血液中のシロリムス濃度を図１に示す。図１（ａ）が真皮中のシロリムス濃度であり、図１（ｂ）が血液中のシロリムス濃度である。真皮中のシロリムス濃度は、１２時間後までは低濃度で一定であり、２４時間後には検出できないほどになった。血液中濃度は、３時間後には上昇し、６時間後および１２時間後には高濃度になった。 7. Result 7-1. Concentrations of sirolimus in the dermis and blood The concentration of sirolimus in the dermis and the concentration of sirolimus in the blood up to 24 hours after a single dose of the 0.4% sirolimus-containing liniment agent (SLN) of Example 1 was administered. It is shown in FIG. FIG. 1 (a) shows the sirolimus concentration in the dermis, and FIG. 1 (b) shows the sirolimus concentration in the blood. The concentration of sirolimus in the dermis was low and constant until after 12 hours, and became undetectable after 24 hours. Blood levels increased after 3 hours and increased after 6 and 12 hours.
比較製剤の０．８％シロリムスゲル剤（ＳＧ）を外用で単回投与した時の２４時間後までの真皮中のシロリムス濃度と血液中のシロリムス濃度を図２に示す。図２（ａ）が真皮中のシロリムス濃度であり、図２（ｂ）が血液中のシロリムス濃度である。 FIG. 2 shows the sirolimus concentration in the dermis and the sirolimus concentration in the blood up to 24 hours after the single administration of the comparative preparation 0.8% sirolimus gel (SG) for external use. FIG. 2A shows the sirolimus concentration in the dermis, and FIG. 2B shows the sirolimus concentration in the blood.
比較製剤の経口製剤（Ｏｒａｌ　Ｓ）を経口で単回投与した時の２４時間後までの真皮中のシロリムス濃度と血液中のシロリムス濃度を図３に示す。図３（ａ）が真皮中のシロリムス濃度であり、図３（ｂ）が血液中のシロリムス濃度である。真皮中にはほとんど移行せず、血液中の濃度は１時間から３時間後に濃度が高くなった後に低下した。 FIG. 3 shows the sirolimus concentration in the dermis and the sirolimus concentration in the blood up to 24 hours after a single oral administration of the comparative oral preparation (Oral S). FIG. 3 (a) shows the sirolimus concentration in the dermis, and FIG. 3 (b) shows the sirolimus concentration in the blood. It hardly migrated into the dermis, and the concentration in blood decreased after 1 to 3 hours after the concentration increased.
７－２．投与したシロリムスの真皮および血液への送達効率
シロリムス投与後の各時点における、真皮へのシロリムスの送達効率（Ｄｓ）（シロリムス投与量の何％が真皮中に存在するか）および血液へのシロリムスの送達効率（Ｂｓ）（シロリムス投与量の何％が血液中に存在するか）を計算した。計算方法は各試験群のシロリムス濃度の中央値を代表値として使用して下記に示す計算式で概算した。外用薬を塗布した領域（約２．５×３．５ｃｍ)の直下にある真皮組織重量を５０ｍｇ、血液循環量を２ｍｌと見積もった。
Ｄｓ＝５０Ｃｄ／１０Ｓｄ
　Ｄｓ：真皮へのシロリムスの送達効率
　Ｃｄ：真皮シロリムス濃度の中央値（ｎｇ／ｍｇ）
　Ｓｄ：投薬シロリムス量（μｇ）
Ｂｓ＝２Ｃｂ／１０Ｓｄ
　Ｂｓ：血液へのシロリムスの送達効率
　Ｃｂ：血液シロリムス濃度の中央値（ｎｇ／ｍＬ）
　Ｓｄ：投薬シロリムス量（μｇ）
結果を図４に示す。本発明の外用剤は、シロリムスが表皮に効率的に浸透した後、真皮に留まらず血中に移行する製剤であり、血液へのシロリムス送達効率は経口製剤と同等であることが示された。 7-2. Efficiency of delivery of administered sirolimus to the dermis and blood At each time point after administration of sirolimus, efficiency of delivery of sirolimus to the dermis (Ds) (what percentage of the dose of sirolimus is present in the dermis) and sirolimus to the blood Delivery efficiency (Bs) (what percentage of the sirolimus dose is present in the blood) was calculated. The calculation method was estimated by the formula shown below using the median sirolimus concentration of each test group as a representative value. The weight of the dermis tissue directly under the area (about 2.5 x 3.5 cm) to which the external preparation was applied was estimated to be 50 mg, and the blood circulation volume was estimated to be 2 ml.
Ds = 50Cd / 10Sd
Ds: Delivery efficiency of sirolimus to the dermis Cd: Median sirolimus concentration in the dermis (ng / mg)
Sd: Dosing sirolimus amount (μg)
Bs = 2Cb / 10Sd
Bs: Efficiency of delivery of sirolimus to blood Cb: Median blood sirolimus concentration (ng / mL)
Sd: Dosing sirolimus amount (μg)
The results are shown in FIG. The external preparation of the present invention is a preparation in which sirolimus efficiently permeates the epidermis and then transfers to the blood, not only in the dermis, and it has been shown that the efficiency of delivering sirolimus to the blood is equivalent to that of the oral preparation.
以上の結果より、本発明の外用剤は、血液中に効率よく送達可能な製剤であることが明らかになった。 From the above results, it was clarified that the external preparation of the present invention is a preparation that can be efficiently delivered into blood.
本発明の外用剤は、投与技術を必要とせず、注射のような痛みを伴わず、乳児や嚥下能の弱った高齢者にも誤嚥を引き起こすことも無く、シロリムスをまたはシロリムス誘導体を血中に投与でき、今後の医療に大いに貢献するものである。 The external preparation of the present invention does not require administration technique, does not cause pain like injection, does not cause aspiration even in infants and elderly people with weak swallowing ability, and sirolimus or sirolimus derivative in blood. It can be administered to the patient and will greatly contribute to future medical care.

Claims (7)

  1. シロリムスおよびシロリムス誘導体からなる群より選択される少なくとも１つと植物油を含有する外用剤。 An external preparation containing at least one selected from the group consisting of sirolimus and sirolimus derivatives and vegetable oil.
  2. 前記植物油が食用油である、請求項１に記載の外用剤。 The external preparation according to claim 1, wherein the vegetable oil is an edible oil.
  3. 前記食用油が、ゴマ油、オリブ油、ダイズ油およびツバキ油からなる群から選択される少なくとも１つである、請求項２に記載の外用剤。 The external preparation according to claim 2, wherein the edible oil is at least one selected from the group consisting of sesame oil, olive oil, soybean oil and camellia oil.
  4. 前記食用油がゴマ油である、請求項３に記載の外用剤。 The external preparation according to claim 3, wherein the cooking oil is sesame oil.
  5. さらに、Ｃ２～Ｃ４のアルカノールを含有する、請求項１～４のいずれか１に記載の外用剤。 Further, containing alkanols C 2 ~ C 4, external preparation according to any one of claims 1-4.
  6. 前記シロリムスおよびシロリムス誘導体からなる群より選択される少なくとも１つが血液中に移行する、請求項１～５のいずれか１に記載の外用剤。 The external preparation according to any one of claims 1 to 5, wherein at least one selected from the group consisting of sirolimus and sirolimus derivatives is transferred into blood.
  7. 前記シロリムス誘導体が、エベロリムス、テムシロリムス、リダフォロリムスおよびゾタロリムスからなる群から選択される少なくとも１つである、請求項１～６のいずれか１に記載の外用剤。 The external preparation according to any one of claims 1 to 6, wherein the sirolimus derivative is at least one selected from the group consisting of everolimus, temsirolimus, lidaphorolimus and zotarolimus.
PCT/JP2020/006722 2019-03-08 2020-02-20 Systemically-acting external preparation containing sirolimus or derivative thereof WO2020184131A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2019042981A JP2022065224A (en) 2019-03-08 2019-03-08 Transdermal-absorbable external preparation
JP2019-042981 2019-03-08

Publications (1)

Publication Number Publication Date
WO2020184131A1 true WO2020184131A1 (en) 2020-09-17

Family

ID=72427600

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2020/006722 WO2020184131A1 (en) 2019-03-08 2020-02-20 Systemically-acting external preparation containing sirolimus or derivative thereof

Country Status (2)

Country Link
JP (1) JP2022065224A (en)
WO (1) WO2020184131A1 (en)

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1919194A (en) * 2006-08-17 2007-02-28 刘瑜玲 Liquid composition of sirolimus
WO2012105521A1 (en) * 2011-01-31 2012-08-09 国立大学法人大阪大学 Externally-used drug for treating skin disorder and method for producing same
CN105663027A (en) * 2016-04-01 2016-06-15 中国人民解放军广州军区武汉总医院 External preparation containing sirolimus as well as preparation method and application thereof
WO2016114216A1 (en) * 2015-01-13 2016-07-21 テルモ株式会社 Biodegradable stent

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1919194A (en) * 2006-08-17 2007-02-28 刘瑜玲 Liquid composition of sirolimus
WO2012105521A1 (en) * 2011-01-31 2012-08-09 国立大学法人大阪大学 Externally-used drug for treating skin disorder and method for producing same
WO2016114216A1 (en) * 2015-01-13 2016-07-21 テルモ株式会社 Biodegradable stent
CN105663027A (en) * 2016-04-01 2016-06-15 中国人民解放军广州军区武汉总医院 External preparation containing sirolimus as well as preparation method and application thereof

Also Published As

Publication number Publication date
JP2022065224A (en) 2022-04-27

Similar Documents

Publication Publication Date Title
Kunzi‐Rapp Topical propranolol therapy for infantile hemangiomas
CN105663027B (en) Sirolimus external preparation, preparation method and the usage
US20210308096A1 (en) Composition for preventing or treating pruritus
US7442690B2 (en) Topical treatment for psoriasis
US7655659B2 (en) Biomimetic of Evodia rutaecarpa fruit extract for amelioration of inflammation
CN105285986A (en) Health food or pharmaceutical composition comprising chestnut shell extract
US20180318398A1 (en) Methods and compositions using ampk activators for pharmacological prevention of chronic pain
JP6556720B2 (en) Daphne Laurera extract in the treatment of dermatoses
WO2020184128A1 (en) Locally-applied external preparation containing sirolimus or derivative thereof
WO2020184131A1 (en) Systemically-acting external preparation containing sirolimus or derivative thereof
CN107982208A (en) A kind of Metformin hydrochloride cream and preparation method thereof
JP2905210B2 (en) Transdermal and transmucosal absorption enhancers and transdermal and transmucosal preparations
US9259451B2 (en) Use of alcohol extract of longan seeds
US20240197803A1 (en) Use of mangosteen fruit shell extract in the preparation of a medicament for treating psoriasis
CN113456594B (en) Method for preparing liposome containing glycyrrhiza inflata extract and madecassoside
CN110088073B (en) Methods and compositions for treating atopic dermatitis
AU2020227426A1 (en) External preparation for vascular abnormality treatment
EP2902034A1 (en) Use of longan seed alcohol extract
JPH06239757A (en) Antiallergic agent
KR102282246B1 (en) Composition for preventing, alleviating, or treating atopic dermatitis
KR102248513B1 (en) A composition for preventing and treating athlete&#39;s foot
US12029771B2 (en) Neem for treatment of RLS
US20210030698A1 (en) Gel containing zerumbone from bitter ginger (zingiber zerumbet) for curative treatment of diabetic ulcers
KR20240032428A (en) Composition for preventing and treating psoriasis comprising extracts of ficus carica
TW202319063A (en) Use of mangosteen fruit shell extract in the preparation of a medicament for treating psoriasis

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: 20769500

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: 20769500

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: JP