CN105147533B

CN105147533B - Bakuchiol composition for treating postinflammatory hyperpigmentation

Info

Publication number: CN105147533B
Application number: CN201510390680.7A
Authority: CN
Inventors: 洪梅芬; 贾琦; 利迪亚·阿尔法洛·布朗奈尔
Original assignee: Unigen Corp
Current assignee: Unigen Corp
Priority date: 2011-02-02
Filing date: 2012-02-02
Publication date: 2018-07-31
Anticipated expiration: 2032-02-02
Also published as: AU2012212153B9; CA3128715A1; WO2012105990A1; CN103458882A; CA3128718A1; KR20200108120A; CA3128711A1; MX350719B; KR102156731B1; KR102642352B1; KR20230058727A; CA2826262C; BR112013019798A2; KR102525516B1; AU2012212153B2; KR20190025740A; HK1215792A1; AU2012212153A1; KR101954118B1; KR20140008364A

Abstract

The method for treating hyperpigmentation is disclosed, the treatment of postinflammatory hyperpigmentation (PIH) is included.Disclosed method includes giving the composition that mammal includes the Bakuchiol that there is no furocoumarin.Also disclose the preparation method of composition and they comprising Bakuchiol.

Description

Bakuchiol composition for treating postinflammatory hyperpigmentation

Citation of related applications

The application requires the US2011/026594 PCT submitted on March 1st, 2011 according to 35U.S.C. § 119 (e) The equity of application and No. 61/438,890 U.S. Provisional Patent Application submitted for 2 days 2 months in 2011.

Background

Technical field

Purposes this invention relates generally to bakuchiol composition and its for treating postinflammatory hyperpigmentation.

Description of Related Art

Postinflammatory hyperpigmentation (PIH) is heavy with increased melanin genesis and the related rare skin pigment of deposition Morbid state.PIH is further characterized in that due to oxidative stress and the mediator from inflammation and immune response and cell factor Invade the apoptosis of the melanocyte generated.Melanocyte deposition (that is, hyperpigmentation) occurs except epidermis level, with aobvious The melanocyte of work is discharged into papillary dermal layer and is captured by big immunocyte.These unique histologic characteristics of PIH are to making Many difficulties are generated with traditional medicament treatment PIH.

The common treatment of PIH concentrate on by using corticosteroid and controlled inflammation using bright protective agent prevent into The pigment of one step develops.The Chemical peeling compound of such as salicylic acid and glycolic also be used to promote skin renewal function simultaneously For removing or reducing pigmentation.Local vitamin A acid also has been used to treatment PIH, but such methods are observing notable effect Up to 40 weeks are needed before benefit.

The tyrosinase inhibitor or skin whitener of such as quinhydrones, azelaic acid, kojic acid and Radix Glycyrrhizae extract have also been used In treatment PIH.Significant drawback using conventional transdermal brightening agent or tyrosinase inhibitor is the normal skin at the neighbouring positions PIH The extensive of skin is faded.The effect makes the color of background skin die down and keeps the positions PIH more prominent.Therefore, on the positions PIH These medicaments must very carefully be applied.In addition, tyrosinase inhibitor is only calm to epidermal pigment excessively effective, this is Because epidermis is the position that melanocyte is synthesized by tyrosinase.Due to after inflammation pigmentation skin deep layer (for example, breast Prominent skin corium) in, therefore the quinhydrones drug for needing continuous 6 months or more before the visible change for observing density bullet Using.Finally, quinhydrones type skin whitener or tyrosinase inhibitor are related to side effect, including skin irritatin, drying, The induction of teratogenesis and leucoderma and cutaneum carcinoma.

The postinflammatory hyperpigmentation energy loses from such as acne, atopic dermatitis, allergic contact dermatitis, pigment Prohibit disease, lichen planus, lupus erythematosus, the endogenous inflammatory skin disorders of morphoea.Other inducements of PIH include such as mechanical Sexual trauma, ionization and Non-ionizing radiation, burn, laser therapy and skin infection exogenous inflammation sexual stimulus.Currently used for above-mentioned The therapeutic agent of skin disorder is invalid for prevention, alleviation, reduction or treatment PIH.For example, be often used such as vitamin A acid, The anti-inflammatory agent treatment of COX inhibitor (for example, salicylic acid), nonsteroidal anti-inflammatory drug (NSAIDs), antiseptic or hormone medicine Above-mentioned skin disorder, but these treatment have been demonstrated it is invalid to PIH.

Although having been achieved with remarkable break-throughs in the field, the field still there is an urgent need for be used to prevent, alleviate, reduce or The method for treating hyperpigmentation.For example, it is desired to the method for treating postinflammatory hyperpigmentation.The present invention realizes These need and provide more associated advantages.

It summarizes

Generally, the present invention relates to the methods for preventing, alleviating, reduce or treat hyperpigmentation.Excessive color Plain calmness can be the result of the morbid state from inflammatory skin disease state.For example, one embodiment of the invention is to use In the method for prevention, alleviation, reduction or treatment postinflammatory hyperpigmentation (PIH).This PIH may originate from many skin diseases Disease, including acne.The method includes give mammalian effective amount comprising Bakuchiol and less than total furan of 500ppm It mutters the composition of cumarin impurity.

With other skin whiteners on the contrary, bakuchiol composition disclosed herein is not tyrosinase inhibitor.Cause This, disclosed composition particularly decolourizes at the positions PIH and for treating in deep skin (for example, papillary dermal layer) Hyperpigmentation.Therefore, presently disclosed compared with being used to treat the method for hyperpigmentation and/or PIH in the past Method includes some advantages.

Therefore, an embodiment of the disclosure is related to for preventing, alleviating, reduce or treat by being originated from inflammatory cutaneous The method of hyperpigmentation caused by the morbid state of illness, the method includes give mammalian effective amount to include Bakuchiol or its pharmaceutically acceptable salt and pharmaceutically acceptable carrier and miscellaneous less than the total furanocoumarins of 500ppm The composition of matter.

In some embodiments, the morbid state is postinflammatory hyperpigmentation.In other embodiments, The composition includes the total furanocoumarins impurity less than 100ppm.In other embodiments, the furocoumarin is miscellaneous Matter includes psoralen, Isopsoralen or combinations thereof.In some embodiments, it is compareed relative to kojic acid, the composition Tyrosinase inhibitory activity is not showed.

In other embodiments, Bakuchiol is chemical synthesis or is detached from plant.For example, in some implementations In scheme, Bakuchiol is detached from plant.In some other embodiment, the plant comes from plant psoralea corylifolia Belong to (Psoralea genus), such as psoralea corylifolia (Psoralea corylifolia L.) (pulse family) or Psoralea Glandulosa L. (Papilionaceae).

In other embodiments, Bakuchiol is from seed, stem, bark, branch, stem tuber, root, root skin, young shoot, root It is detached in stem, flower or other reproductive organs, leaf or other aerial parts or combinations thereof.

In some of the other embodiments, postinflammatory hyperpigmentation (PIH) is originated from acne, atopic dermatitis, allergy Property contact dermatitis, bloch-Siemens syndrome, lichen planus, lupus erythematosus, morphoea, mechanical trauma, ionization or unionized spoke It penetrates, burn, laser or drug therapy, skin infection or combinations thereof.For example, in certain aspects, postinflammatory hyperpigmentation (PIH) it is originated from acne.

In other embodiments, the composition includes the Bakuchiol and medicine of 0.001% to 99.9% total weight , dermatology or the acceptable carrier of beauty.For example, in certain aspects, the composition includes 0.1% to 2.0% total The Bakuchiol of the Bakuchiol of weight, the Bakuchiol of 1.0% total weight or 0.5% total weight.

In other embodiments, the acceptable carrier of dermatology includes nonsticking gauze, bandage, cotton swab, wiping Cloth, plaster, mask or protective agent.In some other embodiments, the acceptable carrier of beauty includes detergent or antibacterial Agent.

In certain aspects, the composition is prepared for local administration.For example, in certain aspects, the composition Also include cream, lotion, ointment, gelling agent, emulsion, liquid, paste, soap, powder agent or combinations thereof.

In other embodiments, the composition also includes adjuvant, skin penetration enhancer or liposome.Other In embodiment, the adjuvant includes 'alpha '-hydroxy acids, salicylic acid, linolenic acid, vitamin A acid, benzoyl peroxide, Sodium Sulfacetamide Sodium, clindamycin, erythromycin, dapsone, tetracycline, doxycycline, minocycline, zinc, estrogen or derivatives thereof resist It is androgen, sulphur, steroids, cortisone, tazarotene, curcumin extract, Arabic gum extract, radix scutellariae extract, green Tea extract, Grape Seed Extract or combinations thereof.

In some embodiments, the composition is prepared with capsule form, for example, controlled release capsule.In other embodiment party In case, by aerosol, by suppository, skin, intramuscular injection or intravenous injection administer locally to the composition.

In certain aspects, the method prevents hyperpigmentation.In other aspects, the method is alleviated excessive Pigmentation.In other aspects, the method reduces hyperpigmentation.In other aspects, the method treatment is excessive Pigmentation.

In other embodiments, hyperpigmentation occurs in the deep layer of skin, for example, the mastoid process in skin is true In cortex.In other embodiments, the method further includes reducing superoxide anion.In some other embodiment party In case, the method further includes reducing melanogen to generate.In other embodiments, the method further includes that reduction melanocyte is thin Born of the same parents are proliferated.In other embodiments, the method further includes preventing melanocyte apoptosis.

In some of the other embodiments, mammal is behaved.In some other embodiments, mammal needs Prevent, alleviate, reduce or treat the hyperpigmentation caused by the morbid state from inflammatory skin disorders.For example, feeding Newborn animal may need to treat PIH.

In another embodiment, this disclosure relates to be generated for reducing melanogen, reduce melanocyte proliferation or prevention The method of melanocyte apoptosis, the method includes giving being connect comprising Bakuchiol or its drug for mammalian effective amount The composition of the salt and pharmaceutically acceptable carrier received and the total furanocoumarins impurity less than 500 ppm.It is other at some Embodiment in, the method further include reduce superoxide anion.

In other embodiments, the composition includes the total furanocoumarins impurity less than 100ppm.In other realities It applies in scheme, furanocoumarin impurities include psoralen, Isopsoralen or combinations thereof.In some embodiments, relatively The composition, which is compareed, in kojic acid does not show tyrosinase inhibitory activity.

In other embodiments, Bakuchiol is chemical synthesis or separation from plant.For example, in some implementations In scheme, Bakuchiol is detached from plant.In some of the other embodiments, Psoralea of the plant from plant (Psoralea genus), such as psoralea corylifolia (Psoralea corylifolia L.) (pulse family) or Psoralea Glandulosa L. (Papilionaceae).

In some embodiments, melanogen generates, melanocyte proliferation or melanocyte apoptosis are that pigment is heavy after inflammation The result of excessive (PIH).In some other embodiments, postinflammatory hyperpigmentation (PIH) is originated from acne, spy Answering property dermatitis, allergic contact dermatitis, bloch-Siemens syndrome, lichen planus, lupus erythematosus, morphoea, mechanical trauma, electricity From or Non-ionizing radiation, burn, laser or drug therapy, skin infection or combinations thereof.For example, in certain aspects, inflammation Hyperpigmentation (PIH) is originated from acne afterwards.

In certain aspects, the method prevents hyperpigmentation.In other aspects, the method is alleviated excessive Pigmentation.In other aspects, the method reduces hyperpigmentation.In other aspects, the method treatment is excessive Pigmentation.In some embodiments, hyperpigmentation occurs in the deep layer of skin, for example, the mastoid process in skin is true In cortex.

In some other embodiments, the method reduces melanogen and generates.In other embodiments, described Method reduces melanocyte proliferation.In other embodiments, the method prevents melanocyte apoptosis.

In some of the other embodiments, mammal is behaved.In some other embodiments, mammal needs It treats and is generated with reducing melanogen, reduces melanocyte proliferation or prevent melanocyte apoptosis.

In other embodiments, the composition also includes salicylic acid or its pharmaceutically acceptable salt.

Other embodiments of the disclosure are related to the method for treating inflammatory or non-inflammatory lesions, and the method includes giving Mammalian effective amount comprising Bakuchiol or its pharmaceutically acceptable salt and salicylic acid or its pharmaceutically acceptable salt and The composition of pharmaceutically acceptable carrier.For example, in some embodiments, lesion includes inflammatory acne lesions.In other realities It applies in scheme, the method treats inflammatory and non-inflammatory lesions.

In some of the other embodiments in front, mammal is behaved.In some other embodiments, lactation Animal needs to treat inflammatory or non-inflammatory lesions.

In other embodiments, the present invention include comprising Bakuchiol or its pharmaceutically acceptable salt and salicylic acid or The composition of pharmaceutically acceptable salt and pharmaceutically acceptable carrier.In some embodiments, the composition is prepared to use In local administration.

It will be evident based on the following detailed description these and other aspects of the invention is referred to.

Brief description

In the accompanying drawings, identical reference number indicates similar element.The size and relative position of element need not in attached drawing Some in drawn to scale and these elements are arbitrarily enlarged and are placed to improve attached drawing legibility.In addition, as drawn The special shape of element is not intended to transmit any information about special elements true form, and is only selected for attached It is readily identified in figure.

Fig. 1 describes the chromatogram of Bakuchiol, psoralen and isopsorapen standard items.

The chromatogram of bakuchiol composition before and after Fig. 2 display hydrolysis.

Fig. 3 provides the data for the strong anti-oxidation property for showing bakuchiol composition.

Fig. 4 is the chart of the tyrosinase inhibitory activity of bakuchiol composition and kojic acid.

Fig. 5 shows the variation of the PIH severity of individual test individuals.

Fig. 6 indicates the chart of the percentage variation of the facial area of the PIH infection of individual test individuals.

Fig. 7 shows the average percent variation of the PIH and PIH severity of five test individuals.

In each interview, the average rank level of PIH and PIH severity reduces compared with benchmark for Fig. 8 descriptions.

Fig. 9 shows photo of two research participants under different time intervals.

Detailed description

In the following description, some details are illustrated to provide the thorough understanding of each embodiment.However, ability Field technique personnel understand can implement the present invention under without these details.In other cases, it is not shown or described in detail Well known structures are to avoid the unnecessary vague description of embodiment.Unless the context requires otherwise, below specification and power Profit require in, word " including (comprise) " and its variant, for example, " including (comprises) " and " including (comprising) " it is interpreted open, inclusive meaning, that is, for example " include but not limited to ".In addition, provided herein Title is not only for for the sake of convenience and indicating that the range or meaning of claimed invention.

In the present specification, to " embodiment (one embodiment) " or " embodiment (an Embodiment reference) " refers to that a particular feature, structure, or characteristic described about embodiment is included at least one reality It applies in scheme.Therefore, phrase " (the in one in one embodiment in each position of this specification Embodiment) " or the appearance of " (in an embodiment) in embodiments " need not be all about identical embodiment party Case.In addition, a particular feature, structure, or characteristic can be combined in any suitable manner in one or more embodiments. In addition, as used in this specification and the appended claims, singulative " a ", " an " and " the " includes plural reference Object, except in addition non-content clearly provides.It is also to be noted that except in addition non-content clearly provides, usual term "or" Use include "and/or" meaning.

Definition

As used herein and unless context dictates otherwise, following term has meaning as defined in following article.

As used herein, " Bakuchiol " refers to the compound for having following formula：

Wherein benzyl double bond can be cis or trans.As used herein, Bakuchiol include pharmaceutically acceptable salt and The tautomer of Bakuchiol.It is also included in this definition with the relevant oxybenzene compound of Bakuchiol structure.

“Bakutrol^TM" be the composition comprising Bakuchiol and can also additionally comprise from Psoralea (Psoralea) aliphatic acid extracted in plant.

" UP256 " refers to 0.5% (wt/wt) preparation of Bakuchiol.

" preventing (Preventing) ", " preventing (prevention) " in the context of published method and " prevention (prevent) " all refer to prevent or stop such as PIH special disease state generation prevention method.

" alleviating (Alleviating) ", " alleviating (alleviation) " in the context of published method and " alleviation (alleviate) " all refer to alleviation or the influence of the special disease state of mitigation such as PIH or symptom.

" reducing (Reducing) ", " reducing (reduction) " in the context of published method and " reduction (reduce) " all refer to reduce such as PIH special disease state influence or symptom.

" treatment (Treating) ", " treatment (treatment) " in the context of published method and " treatment (treat) " all refer to be intended to improve such as PIH special disease state symptom reduction or stop its generation technology or Method.

" impurity " includes any substance not expected in bakuchiol composition, is typically derived from from natural origin The Bakuchiol of middle separation.Term impurity includes but not limited to furocoumarin compound, the furocoumarin compound packet Include but be not limited to psoralen, Isopsoralen and other cumarin type dopants.Impurity also refers to from these groups of acquisition Close the impurity of the synthetic method of object.

" treatment " includes treatment and/or prevents.When in use, treatment refers to people and other animals.

" pharmacy, beauty or treatment effective dose or amount " refers to the agent for being enough to induce desired biology or function result Amount is horizontal.The result can be to alleviate disease, the sign of skin disease state, symptom or inducement or desired biosystem Any other change.

" placebo " refers to the expectation of the sign, symptom or inducement that are enough that usable inert matter is induced to alleviate disease Biology drug treatment effective dose or amount administration substitute.

" receptor (host) " or " individual " or " patient " be the living individuals for being given compositions described herein, people or Animal.Therefore, compositions described herein can be used for animal doctor and people is applied and should not be explained that term " is suffered from a manner of limitation Person " or " individual " or " receptor ".In the case of veterinary application, it can be given as described below according to the weight of animal described Dosage range.

As described above, an embodiment of the disclosure is related to using comprising there is no furanocoumarin impurities The composition of Bakuchiol is for preventing, alleviating, reducing or treating the mistake caused by the morbid state from inflammatory skin disorders Spend pigmentation.For example, disclosed method is suitable for treatment postinflammatory hyperpigmentation (PIH).In some embodiments In, PIH may originate from acne.Disclosed method has shown to be originated from such as acne, idiocrasy skin in prevention, alleviation, reduction and treatment Inflammation, allergic contact dermatitis, bloch-Siemens syndrome, lichen planus, lupus erythematosus, morphoea skin disorder inflammation after color It is plain calm excessive；With color after the inflammation caused by mechanical trauma, ionization and Non-ionizing radiation, burn, laser and drug therapy It is plain calm excessive, and by using synthesis Bakuchiol or not Psoralea (psoralea) extraction of furocoumarin People's clinical efficacy in the skin infection of object bakuchiol composition.Based on refer to subsequent description, the disclosure these and its Its aspect and each embodiment will be apparent.

A. bakuchiol composition

In one embodiment, present disclose provides the compositions comprising Bakuchiol, and the Bakuchiol is substantially There is no impurity, especially furanocoumarin impurities.The composition is being also known as Bakutrol herein^TM.In some embodiments In, such as in document, ((organic chemistry is logical by Hongli Chen and Yuanchao Li, Letters in Organic Chemistry News), 2008,5,467-469) shown in by organic synthesis from simple compounds or from plant obtain the composition. In some embodiments, bakuchiol composition is detached from plant.The plant source of Bakuchiol includes plant The section of section, the plant includes but not limited to pulse family (Luguminosae), Papilionaceae (Papilionaceae), Lauraceae (Lauraceae) and Magnoliaceae (Magnoliaceae) and plant category, it includes but not limited to Psoralea that the plant, which belongs to, (Psorlea), Sassafras (Sassafras), Magnolia (Magnolia) and [WTBX (Astractylodes).For example, mending Bone fat phenol composition can be from psoralea corylifolia (Psoralea corylifolia L.) (pulse family) or Psoralea glandulosa L. it is detached in (Papilionaceae).One or more unitary parts that can be from entire plant or from plant include but not limited to kind Son, stem, bark, branch, stem tuber, root, root skin, young shoot, rhizome, flower or other reproductive organs, leaf or other aerial parts Or combinations thereof in obtain the composition.Method for detaching Bakuchiol from plant may include solvent extraction, overcritical Fluid extraction, distillation, physical squeezing or combinations thereof.

Bakuchiol, structure illustrate below, are the benzene with a hydroxyl and aliphatic unsaturated hydrocarbon in aromatic ring Phenolic compounds.Although being indicated with trans forms in having structure, the benzyl double bond of Bakuchiol is also likely to be cis-.

The amount (that is, weight percent (w/w%)) of Bakuchiol depends on extracting process and thick in pure plant extract The degree of purification of extract.In one embodiment, as shown in table 2, in extract Bakuchiol amount be 13.7% to 29.1%.In other embodiments, in extract the amount of Bakuchiol be at least 30%, at least 35%, at least 40%, extremely Few 45%, at least 50%, at least 60%, at least 70%, at least 80% or at least 90%.In some embodiments, it extracts The amount of Bakuchiol is 100% in object.In some other embodiment, the amount of Bakuchiol is not less than in the composition 60%.Embodiment 6-8 provides the example of the extract of the Bakuchiol comprising each amount.

Although Bakuchiol is living for preventing and treating the biology with very big potentiality of various diseases and morbid state Property natural products, but exist many restrictions related with the use of the compound.Some limitations include that it is low in natural source The presence of common existing toxic component in concentration and Bakuchiol source.Impurity is with Psoralen present in bakuchiol composition Fat phenol source and change.For example, psoralen, also known as furocoumarin, are planted at Psoralea (Psoralea genus) Naturally occurring secondary metabolites and also exist in many fruits and vegetables in object (a kind of Bakuchiol source).Often find Example with Bakuchiol existing furocoumarin jointly includes psoralen and isopsorapen.

Many health risks are known as with the plant of processing, topical application and intake containing psoralen and synthesis psoralea corylifolia It closes.Known psoralen is phototoxicity agent, increases skin to the sensibility of ultraviolet radiation and promotes cutaneum carcinoma (Epstein (1999)Med.Surg.18(4)：274-284).Psoralen is proved growth inhibition (Diawara et al. in induced rat (1997)Cancer Lett.114(1-2):159-160).The property of thick extract from Psoralea (Psoralea) plant The destruction of gland toxicity and hypothalamic pituitary gonadal axis, which has, directly contacts (Takizawa et al. (2002) J.Toxicological Sciences (toxicology Scientific Periodicals) 27 (2):97-105).It is taken orally in the diet of female rats and gives psoralen, Buddhist Hand lactone (5-MOP) and xanthotoxin (8-methoxypsoralen) reduce birth in a dose-dependent manner Rate, the quantity of transplantation site, doggie, corpus luteum, full and empty uterus weight and circulating estrogen level (Diawara et al. (1999) J. Biochem.Molecular Toxicology (biochemical molecule toxicology periodical) 13 (3/4):195-203).Psoralen Fat element also has been found the mRNAs of induction liver enzyme CYP1A1 and UGT1A6, shows the metabolism that estrogen is improved by psoralen Reduction (Diawara et al. (June in May, 2003 -) of interpretable genotoxicity and folliculus ovarii function and ovulation observation Pediatr Pathol Mol Med.22(3):247-58.).Due to furocoumarin toxicity, it is therefore important that from intention Psoralen and different benefit are removed in bakuchiol composition for treating postinflammatory hyperpigmentation or Other diseases state Bone fat element.

Psoralen and isopsorapen accounts for the dry weight of the Psoralea seed of about 0.1%-2% and in solvent or super The weight of about 1%-20% is accounted in supercritical fluid extraction object.Solvent extraction or supercritical fluid extraction, distillation, physics can be passed through Squeezing or the combination of above-mentioned extracting process obtain the thick extract from Psoralea (Psoralea genus) plant.It can be logical Cross chromatographic isolation, demixing of solvents (indian patent discloses #00570/KOL/2005), distillation, recrystallization and other Wet chemicals and Physical method obtains the bakuchiol composition of enrichment.Disclosed No. 2006/0251749 U.S. Patent application, with its entirety Content is incorporated herein by reference, and discloses the subsequent hydroxylating of solvent extraction to decompose furocoumarin ring and obtain substantially There is no the bakuchiol composition of the enrichment of furanocoumarin impurities (for example, the furans less than 500ppm or less than 100ppm is fragrant Legumin impurity).Disclosed method include from plant source extract compounds, using aqueous slkali hydrolyze thick extract under heating With by include but not limited to column chromatography, extraction subsequent crystallisation, demixing of solvents, recrystallization and combinations thereof method purifying step Suddenly.Applicants have discovered that the Psoralea of this Bakuchiol enrichment that there is no furanocoumarin impurities (psoralea) composition of extract can be used to preventing, alleviate, reduce or treating hyperpigmentation.For example, disclosed benefit Bone fat phenol composition is effective to prevention, alleviation, reduction or treatment postinflammatory hyperpigmentation (PIH).

Present disclosure also relates to for detaching and purifying Bakuchiol thick composition and the related compounds that are obtained from natural source The method of object.Method for detaching and purifying these compositions include from plant source extract compounds, using aqueous slkali water Solve thick extract and the side by including but not limited to column chromatography, extraction subsequent crystallisation, demixing of solvents, recrystallization and combinations thereof The step of method purifies.The thick extract purified in this way there is no the furans of such as psoralen and isopsorapen Cumarin impurity.Therefore, potential phototoxicity related with these compounds, local irritation, carcinogenicity and genotoxicity are basic On be eliminated.

In some embodiments, disclosed composition include less than 500ppm, be less than 250 ppm, be less than 100ppm or Total furanocoumarins impurity less than 50ppm.Furocoumarin can be measured by any method known to those skilled in the art The concentration of impurity.For example, in one embodiment, furocoumarin content can be measured by HPLC.

As described in example 2, under two groups of extraction conditions using six kinds of different organic solvent systems evaluations from The effect of Bakuchiol extraction extracted in plant source.As a result it is illustrated in table 2.Reference table 2, it can be seen that many can be used Organic solvent and/or a combination thereof extract Bakuchiol from Psoralea (Psoralea) plant.Psoralea corylifolia in each extract The amount of phenol is 13.7% to 29.1% weight ratio.Other extracting process include but not limited to CO₂Supercritical fluid extraction and water Distillation.The squeezing diffusate of fresh plant part from such as seed can be used for obtaining Bakuchiol group from natural source Close object.

The effect that thick Bakuchiol extract is purified by column chromatography is shown in embodiment 3 and table 3.Especially have rated Eight kinds of different type resins detach the ability of Bakuchiol from furanocoumarin impurities.Silica column and CG-161 resins two Person shows satisfactory separation.However, the thick plant extract of column chromatography for separation is not usually economically feasible at industrial scale , because it needs expensive equipment and reagent and experienced person.Since the complexity of thick plant extract causes The low-down load capacity of these samples also make commercial scale column chromatography difficult.

Embodiment 4 describes the economic means for detaching Bakuchiol from furanocoumarin impurities.The method packet Include the composition for including furanocoumarin impurities using alkali process.As illustrated in by following scheme 1, in order to illustrate using NaOH, the lactonic ring of furocoumarin is opened using alkali heating, thus converts them into corresponding carboxylate.Then, energy By a variety of methods these salt are easily separated from the remainder of mixture.Disclosed method, which allows to prepare, there is no The bakuchiol composition of furanocoumarin impurities (for example, being less than 500ppm).Standard is used under not disclosed hydrolysis Chromatographic technique cannot obtain the bakuchiol composition that so height is pure.

The hydrolysis of 1. furocoumarin of reaction scheme

Aqueous slkali may include any alkali that can open lactonic ring, including but not limited to sodium hydroxide, potassium hydroxide, hydrogen Calcium oxide, lithium hydroxide or combinations thereof.Solution can have various concentration and pH values to be converted into hydrochlorate to the maximum extent.It can also Reaction mixture is heated at different temperatures and pressures so that reaction rate, efficiency and yield maximize.

Reaction process then can be HPLC to ensure that furocoumarin is fully converted to their corresponding carboxylates.Hydrolysis Before and after the HPLC chromatogram of thick composition illustrate in fig. 2.Reaction after the completion of (such as measured by HPLC ), a variety of methods can be used to handle reaction solution, including but not limited to column chromatography, crystallization, demixing of solvents, precipitation, solvent is washed Wash or combinations thereof.The organic solvent that can be used for demixing of solvents includes but not limited to petroleum ether, ethyl acetate, ether, hexane, chlorine Imitative, propyl alcohol, butanol and dichloromethane and the immiscible organic solvent of other water.

The thick extract purified in this way there is no the furocoumarin of such as psoralen and isopsorapen Impurity.For example, the extract of purifying may include being less than 500ppm, be less than 250ppm, be less than 100ppm or even less than 50ppm Furanocoumarin impurities.In addition, these pure colors without furocoumarin, bakuchiol composition of height are light brown Color or red and they about activating agent color and form it is highly stable so that they are especially suitable for preparing, storing And cosmetic applications.

It is the method for the composition for analyzing Bakuchiol that the disclosure is also included, can detect and quantify impurity. In this embodiment, the method for analyzing bakuchiol composition includes analyzing institute by high pressure liquid chromatography (HPLC) The step of stating composition.The quantitative of each component in mixture, which can be realized, by HPLC analyses and additionally provides tracking mends The method of Bakuchiol, psoralen, Isopsoralen and other natural constituents in bone fat category (Psoralea) plant with Guidance extraction, hydrolysis and purification process.The method of the composition of Bakuchiol is analyzed using high pressure liquid chromatography (HPLC) in reality It applies described in example 1 (table 1).

B. bakuchiol composition is used to treat hyperpigmentation

One embodiment of the disclosure is related to using comprising the Bakuchiol that there is no furanocoumarin impurities Composition sinks for preventing, alleviating, reducing or treating the excessive pigment caused by the morbid state from inflammatory skin disorders It.For example, disclosed method includes preventing, alleviate, reduce or treating postinflammatory hyperpigmentation (PIH).In some implementations In scheme, PIH may originate from acne.The disclosure is included in typical beauty medium and also in face cream, moisturizing Bakuchiol composition is prepared in dew (gel lotion) and the other preparations being described in more below.As shown in embodiment Show, the applicant has confirmed bakuchiol composition in prevention, alleviation, reduction or treatment postinflammatory hyperpigmentation (PIH) unexpected people's clinical efficacy in, wherein PIH connect from skin disorder such as acne, atopic dermatitis, anaphylaxis Touch property dermatitis, bloch-Siemens syndrome, lichen planus, lupus erythematosus, morphoea；With by mechanical trauma, ionization and unionized spoke It penetrates, burn, postinflammatory hyperpigmentation and skin infection caused by laser and drug therapy.

Disclosed method includes that there is no furans perfume including it is a effective amount of to give mammal (for example, people patient) The composition of the Bakuchiol of legumin impurity.For example, the composition may include the furanocoumarin impurities less than 500ppm. The composition can include about the Bakuchiol of 0.0001% to about 100%.For example, in some embodiments, the combination Object includes the Bakuchiol of about 0.1% to about 2% or the Bakuchiol of about 0.5% to about 1%.In other examples, described Composition includes about 0.5% or about 1.0% Bakuchiol.In some embodiments, mammal is people, and at it In its embodiment, mammal needs to prevent, alleviate, reduce or treat to be drawn by the morbid state from inflammatory skin disorders The hyperpigmentation risen, such as the mammal may need to treat PIH.

The disclosure shows the unexpected unique biological property of synthesis or natural bakuchiol composition.As implemented Shown in example 5 and table 4, including the Bakutrol compositions of about 57.35% Bakuchiol have unexpectedly high anti-oxidant energy Power, particular against superoxide anion (>69,000 μm of ole TE/g), five kinds of chief active kinds of confrontation, which have, to be in> Total ORAC values of 92,000 μm of ole TE/g.

Superoxides is with chemical molecular formula O₂ ^-Anion.The chronic inflammatory disease state energy of such as acne vulgaris With the superoxide anion that the slave keratinocyte dramatically increased generates, by such as propionibacterium acnes (P.acnes) Gram-positive anaerobic bacterium stimulates (Grange PA. et al., Plos Pathogens (Public science library pathogen) 2009,5 (7) 1-14.).Superoxides be bio-toxicity it is very high and scattered by immune system with kill invasion it is micro- Biology.In phagocyte, superoxides largely generates the oxygen dependence for invading pathogen by enzyme nadph oxidase Kill mechanism.Superoxide anion and other reactive oxygen species in inflammation skin also can induce melanogen generate, melanocyte it is thin Born of the same parents are proliferated and melanocyte apoptosis, are the main pathogenic of postinflammatory hyperpigmentation.Therefore, one of the disclosure Embodiment is to reduce super oxygen by using the composition comprising the Bakuchiol that there is no furanocoumarin impurities Object is come the method alleviating, reduce or treat the hyperpigmentation caused by the morbid state from inflammatory skin disorders.One In a embodiment, morbid state PIH.In another embodiment, present disclose provides reduce melanogen generation or melanocyte Cell Proliferation or the method for inhibiting melanocyte apoptosis, for example, by reducing superoxide anion.The method includes giving Give the composition for including the Bakuchiol that there is no furanocoumarin impurities of mammalian effective amount.In some implementations In scheme, mammal is people, and in other embodiments, and mammal needs to reduce melanogen generation or melanocyte is thin Born of the same parents are proliferated or inhibit melanocyte apoptosis.

As embodiment 6 and Fig. 3 are confirmed, including 77.02% there is no that impurity, especially furocoumarin are miscellaneous The composition of the Bakuchiol of matter shows the protective effect of the oxidative stress to being induced by 4- tetrabutyls phenol (4-TBP).Two It tests under a concentration, is combined by Bakuchiol to carrying out the cytotoxicity of melanocyte for the reactive oxygen species that free 4-TBP is generated Object is protected.Although undesirable by the theoretical constraint, applicants contemplate that derived from synthesis or natural bakuchiol composition Reduction, alleviation, prevention or treatment postinflammatory hyperpigmentation (PIH) unexpected clinical benefit from its solely Special and unexpected neutralization activity oxygen type, the especially ability of superoxide anion, and causing reduced epidermis black Protect melanocyte from oxidative stress under skin spot and/or the Inflammatory disease states of corium melanose.

Other than its unexpected high oxidation resistance, applicants have discovered that disclosed bakuchiol composition is not It is tyrosinase inhibitor.This is with open Bakuchiol as the skin whitener (P1107123 inhibited by tyrosinase Number Japan Patent) other reports it is opposite.This is it has unexpectedly been discovered that guiding the applicant reaches for treating the current of PIH Disclosed method, wherein pigmentation occur in deep skin layer and tyrosinase inhibitor is invalid.It is disclosed No tyrosinase inhibit bakuchiol composition shown in embodiment 7 and Fig. 4.Pure Bakuchiol (100%) and Both enrichment Bakuchiols (77.02%) with the furocoumarin from natural source no more than 100ppm at eight kinds not Inhibit function with there is no tyrosinase under dosage.

Evaluate the safety of the composition comprising Bakuchiol under a concentration of 86.54% and 77.02% Bakuchiol Property.As shown in embodiment 9 and table 6, eyes are not showed based on external and people's clinical test, Bakutrol (UP256) composition Irritation does not have skin irritation to normal or galling skin, does not have skin contact sensitization, no phototoxicity and do not have There is mutagenic toxicity.The topical cream of bakuchiol composition all well-tolerated in owner and in vitro test.

As embodiment 10 is shown, with from slight or moderate acne vulgaris postinflammatory hyperpigmentation (PIH) on individual, furans tonka-bean of the test comprising 77.02% Bakuchiol and less than 100ppm in people's clinical test The natural bakuchiol composition of the seed extraction and enrichment of the slave psoralea corylifolia (Psoralea corylifolia) of element (Bakutrol^TM).Bakuchiol composition is prepared under 0.5% Bakuchiol is used for topical application.In daily topical application After 0.5% Bakutrol emulsifiable pastes, significantly subtracting for postinflammatory hyperpigmentation (PIH) is observed in all five individuals It is few.As shown in figure 5, all five individuals have the reduction of the PIH severity of at least one hierarchy level.Continued office at 8 weeks The improvement (Fig. 6) of the facial area of PIH influence of the portion using realization after 0.5% Bakutrol emulsifiable pastes more than 50%.PIH And average percent and absolute scale the level improvement of both its severity are summarized in figures 7 and 8.Early in using psoralea corylifolia Phenol composition is achieved that the improvement for being more than 40% of both PIH and severity after 4 weeks, or is more than a hierarchy level Reduction.As shown in figure 9, in two individual photos on the skin of face position of influence PIH be greatly decreased it is aobvious and easy See.After topical application Bakutrol emulsifiable pastes, pigment after two individual displays scytitis related with slight and moderate acne The gradually improvement of calm excessively (PIH).

Table 7 (embodiment 10) outlines and the popular acne treatment product phase that includes antiseptic or anti-inflammatory agent or combinations thereof Than using the clinical success of the bakuchiol composition (that is, Bakutrol) of not furocoumarin.Data in table 7 understand It shows that the bakuchiol composition of not furocoumarin not only improves inflammatory and non-inflammatory lesions count, but also significantly improves Hyperpigmentation after scytitis.Lack tyrosinase inhibition activity based on it, PIH, which has benefited from bakuchiol composition, is It is unexpected.

Table 8 (embodiment 11) provides the data of the reduction of display PIH grades (that is, pigmentation level).Data understand Show Bakuchiol than for treating PIH placebo and salicylic acid it is more effective.In addition, applicant have also found that Bakuchiol (or including the composition of the Bakuchiol) is effective for the inflammatory lesion for treating such as acne lesion.Table 9 (embodiment 11) It shows compared with using placebo or salicylic treatment, validity of the Bakuchiol for treatment inflammatory lesion.

Other than including the method treated using the composition comprising Bakuchiol, the present invention includes wherein using packet The embodiment for treating mammal containing Bakuchiol and salicylic composition.For example, applicants have discovered that salicylic acid is to controlling It is effective to treat non-inflammatory lesions, while Bakuchiol is effective to treatment inflammatory lesion.Therefore, one embodiment of the invention relates to And the method for the treatment of inflammatory lesion (for example, acne lesion), include to mend the method includes give mammalian effective amount The composition of bone fat phenol or its pharmaceutically acceptable salt.Another embodiment is related to treating inflammatory and/or non-inflammatory lesions (example Such as, acne lesion) method, the method includes give mammalian effective amount comprising Bakuchiol and salicylic acid (or its Pharmaceutically acceptable salt) composition.Other embodiments include by give mammalian effective amount comprising salicylic acid or The composition of its pharmaceutically acceptable salt treats non-inflammatory lesions.In some embodiments in front, mammal is behaved. In other embodiments, mammal needs to treat the inflammatory and/or non-inflammatory lesions of such as acne.

Other than treating lesion, Bakuchiol and it is salicylic combination to treatment aforementioned diseases state (for example, PIH, Melanogen is reduced to generate, reduce melanocyte proliferation or prevent melanocyte apoptosis etc.) any one of effectively.Therefore, some Embodiment is related to use and is treated comprising Bakuchiol and salicylic composition.Other embodiments include comprising psoralea corylifolia The composition of phenol or its pharmaceutically acceptable salt, salicylic acid or its pharmaceutically acceptable salt and pharmaceutically acceptable carrier.

Preceding method is effective to substantially eliminating inflammatory and/or non-inflammatory lesions.For example, in some embodiments, institute It states method and reduces lesion about 1% to about 99% or about 10% to about 90%.In other embodiments, the method reduces disease Become and is more than 50%.

Bakuchiol and salicylic ratio are not particularly limited and can be based on it is expected by persons skilled in the art As a result it determines.For example, in some embodiments, Bakuchiol and salicylic weight ratio are about 1:100 to about 100:1. In other embodiments, weight ratio is about 10:90, about 20:80, about 30:70, about 40:60, about 50:50, about 60:40, about 70: 30, about 80:20 to about 10:90.The composition can be prepared according to any formula described herein.

C. the preparation of bakuchiol composition

The bakuchiol composition of the disclosure can be prepared by any method known to those skilled in the art.As implemented Shown in example 8 and table 5, the composition of the disclosure can be prepared in the form of pharmacy, beauty or dermatological compositions, and can include Other components, such as pharmacy and/or the acceptable active matter of beauty, excipient, adjuvant, carrier or combinations thereof.Excipient is to use Make the diluent of dermatology and the acceptable prodrug and drug of beauty or the inert substance of medium.The reality of this kind of excipient Example includes but not limited to water, buffer, brine, glycerine, hydrated SiO 2, propylene glycol, aluminium oxide, carrageenan, fibre The plain carboxylic methyl ether of dimension, titanium dioxide, Ringer's solution, glucose solution, mannitol, Hank's solution, preservative and other aqueous Physiological equilibrium salting liquid.The non-aqueous vehicles of such as fixing oil, sesame oil, ethyl oleate or triglycerides also can be used.Its Its useful preparation includes the suspending agent for including such as tackifier of sodium carboxymethylcellulose, sorbierite or glucan.

In embodiment 8, in carbitol or Trivent OCG or polysorbate -20 or pure water or two kinds or The composition of the disclosure is prepared in the combination of a variety of above-mentioned mediums.Excipient can also include micro additive, such as EDTA, disodium DDTA, BHA, BHT, dibasic ammonium citrate, nordihydroguaiaretic acid, propylgallate, sodium gluconate, partially Bisulfite receives, tert-butyl hydroquinone, SnCl₂、H₂O₂With 2,4,5- THBP 2,4,5 trihydroxybutyrophenones, vitamin C, vitamin E, acetic acid dimension Raw element E, phenonip and other substances for improving isotonicity and chemical stability.

The example of the substance of pH for adjusting preparation include sodium hydroxide, sodium carbonate, sodium bicarbonate, sodium triphosphate, Tetrasodium pyrophosphate, lauryl sodium sulfate, calper calcium peroxide, phosphate buffer, bicarbonate buffer, tris buffer solutions, group Or mixtures thereof propylhomoserin, citrate and glycine.The example of flavoring agent includes but not limited to thimerosal, metacresol or adjacent first Phenol, formalin, fruit extract and benzylalcohol.Standard preparation can be liquid or solid, can in suitable liquid such as suspension or It is dissolved in solution for being administered.Therefore, in non-liquid formulation, excipient can include glucose, human serum albumins, Before administration sterile water or brine can be added to it in preservative etc..

In one embodiment, the activation of the different role mechanism of cutaneous pigmentation can be reduced with other targetings It closes object and prepares bakuchiol composition together.This kind of active matter includes but not limited to quinhydrones, single-benzyl ether, ursin (arbuting), deoxyarbutin, p methoxy phenol, N- acetyl group -4-S- cysteamine bases phenol, kojic acid, azelaic acid, ethyl alcohol Acid, gentianic acid, flavonoids, Aloesin, talan and diphenyl ethylene derivatives, Radix Glycyrrhizae extract, black bearberry extract, mulberry Mulberry extract, Aloe Vera Gel, glabridin, vitamin C derivatives, magnesium ascorbyl phosphate, tetrahexydecyl ascorbate, dimension Raw element E derivatives, tranexamic acid and its derivative, the biosimulation object of TGF-B protein, centaurcidin, niacinamide, PAR-2 inhibitor, agglutinin, Neoglycoproteins, resorcinol and its derivative and Nivitol^TM。

In another embodiment, the composition include can with bakuchiol composition act synergistically anti-inflammatory agent and Antiseptic is to reduce infection, the relevant inflammation of infection and accelerate epidermal renewal.This kind of active matter includes but not limited to Alpha-hydroxy Acid, salicylic acid, linolenic acid, vitamin A acid, benzoyl peroxide, Guttae Sulfacetamidi Natrici, clindamycin, erythromycin, dapsone, four Ring element, doxycycline, minocycline, zinc, estrogen and its derivative, antiandrogen, sulphur, steroids, cortisone, he prick Luo Ting, curcumin extract, Arabic gum extract, radix scutellariae extract, Green tea extract and Grape Seed Extract.

In some embodiments, the composition includes adjuvant or carrier.In general, adjuvant is usually to enhance mammal To the substance of the biological respinse of particular bioactive agent.Suitable adjuvant includes but not limited to Fo Shi reagents；Other bacterial cells Wall fraction；Aluminium, calcium, copper, iron, zinc, magnesium, tin class salt；Silica；Crystallite grinds skin agent, polynucleotides；Toxoid；Haemocyanin Matter；Viral capsid proteins；Other bacterial derivation preparations；Interferon；Such as block copolymerization of Heng Teshi Titermax adjuvants Object adjuvant (Vaxcel.TM., Inc.Norcross, Ga.)；Ribi adjuvants (Ribi ImmunoChem Research are purchased from, Inc., Hamilton, Mont.)；With saponin(e and its derivative, such as Quil A (be purchased from Superfos Biosector A/S, Denmark).Carrier is usually the compound for the half-life period for increasing therapeutic combination in treated receptor.Suitable carrier Including but not limited to polymer release-control preparation, biodegradable implant, liposome, Nano capsule, nano particle, bacterium, Virus, oil, esters and glycols.

In other examples, preparing the composition with the controlled release formulation of composition described in slow release to receptor. As used herein, controlled release preparation is included in the bakuchiol composition in controlled release vehicles object.Suitable controlled release vehicles object is this Known to field technology personnel.The example of controlled release preparation is biodegradable (that is, biology is digestible) and includes capsule Agent.

In one embodiment, suitable ointment is generally selected by the total weight based on topical formulations of expectation concentration From 0.001% to 100% effective, non-toxic, amount UP256 (Bakuchiol), 65% to 100% (for example, 75% to 96%) White soft paraffin, 0% to 15% liquid paraffin and the lanolin of 0% to 7% (for example, 3% to 7%) or its spread out Biology or synthesis equivalent composition.In another embodiment, ointment may include polyethylene-liquid paraffin matrix.

In one embodiment, suitable cream by emulsification system together with expectation concentration synthesize and/or from upper UP256 (Bakuchiol) compositions detached in the single plant or various plants of offer are provided.Emulsification system preferably comprise 2% to 10% polyoxyethylene alcohol (for example, being mixture of CetomacrogolTM1000 acquisitions with trade mark), 10% to 25% Stearyl alcohol, 20% to 60% liquid paraffin and 10% to 65% water；Together with one or more preservatives, for example, 0.1% to 1% N, N "-di-2-ethylhexylphosphine oxide [N '-[3- (methylol) -2,5- dioxo -4- imidazolidinyls] urea] are (with entitled miaow urea USNF is obtained), 0.1% to 1% 4-HBA Arrcostab is (such as by the entitled Nipastat of trade mark purchased from Nipa The mixture of Laboratories), 0.01% to 0.1% 4-HBA sodium butyrate is (with the entitled Nipabutyl of trade mark Sodium be purchased from Nipa Laboratories) and 0.1% to 2% Phenoxyethanol.

In one embodiment, suitable gel is limited in crosslinked three-dimensional poly- with high level by wherein liquid phase Semisolid systems composition in polymer matrix.Liquid phase may include water together with desired amount UP256 (Bakuchiol), 0.01% to The 20% such as additive and 0.01% to 10% of glycerine, polyethylene glycol or the water soluble of propylene glycol, preferably 0.5% to 2% thickener, the thickener can be natural products, such as tragacanth, pectin, carrageenan, agar and alginic acid, or Person synthesizes or semi-synthetic compound, such as methylcellulose and carbopol (carbopol)；Together with one or more preservatives, Such as 0.1% to 2% 4-HBA methyl esters (methyl p-hydroxybenzoate) or Phenoxyethanol-difference (differential).In addition suitable preparation by desired amount UP256 (Bakuchiol), together with 70% to 90% poly- second Glycol (for example, the polyethylene glycol ointment agent comprising 40% polyethylene glycol 3350 and 60% polyethylene glycol 400, according to the U.S. Prepared by formulary (USNF)), 5% to 20% water, 0.02% to 0.25% antioxidant (such as Butylated hydroxy first Benzene) and 0.005% to 0.1% chelating agent (such as ethylenediamine tetra-acetic acid (EDTA)) composition.

The term soft paraffin being used above includes cream or ointment off-white color soft paraffin and yellow soft paraffin.Term Lanolin includes natural wool fat and refined wool fat fat.It the derivative of lanolin particularly including has been chemically modified to change Become their lanolin physically or chemically and the synthesis equivalent of lanolin particularly including it is known and for drug and Synthesis or semi-synthetic compound and mixture of the beauty treatment fields as the substitute of lanolin and it may for example be known as lanolin Substitute.

A kind of suitable synthesis equivalent of workable lanolin is the substance that the entitled SoftisanTM of trade mark is obtained, It is referred to as Softisan 649.Softisan 649 is natural purchased from Dynamit Nobel Aktiengesellschaft The glycerine ester of vegetable fatty acid, isostearic acid and aliphatic acid；Its property by H.Hermsdorf in Fette, Seifen, Anstrichmittel, issue number 84 discuss in No.3 (1982), pp.3-6.

Exist as the other materials mentioned above of the ingredient of suitable ointment or cream base and their property It is discussed in standard reference, such as United States Pharmacopeia.Cetomacrogol 1000 has general formula CH₃(CH₂)_m(OCH₂CH₂)_nOH, Wherein m can be that 15 or 17 and n can be 20 to 24.Butylated hydroxytoluene is 2,6- di-t-butyls-paracresol.Nipastat is The mixture of 4-HBA methyl esters, ethyl ester, propyl ester and butyl ester.

Compositions disclosed herein can be prepared by conventional phamaceutical techniques.Thus, for example, can be by high temperature for example At 60 DEG C -70 DEG C by soft paraffin, if there is liquid paraffin and lanolin or derivatives thereof or synthesis equivalent be blended in one It rises and easily prepares above-mentioned composition.Then, the mixture can be cooled to room temperature and in the hydration that mupirocin is added After crystallizing calcium salt, together with corticosteroid and any other ingredient, stirring is fully dispersed to ensure.

Finally, Bakuchiol has the distribution coefficient of log P=6.13.The distribution coefficient of chemical compound provides it Hydrophily/Lipophilic Balance thermodynamics detection, thus provide its potential source biomolecule availability.With 6.13 distribution Coefficient refers to that the compound has high membrane permeability and bioavilability when being prepared in delivery system.In separation The Cutaneous permeation of reactive compound-Bakuchiol in face cream is quantified in vitro test in application on human skin.As a result it shows good Cutaneous permeation and bioavilability.In some embodiments, disclosed composition includes skin penetration enhancer.

D. bakuchiol composition is given

The composition of the disclosure can be given by any method known to those of ordinary skill in the art.For example, can take orally Or administer locally to disclosed composition.Medication includes but not limited to (oral) administration of enteral, parenteral (intravenous injection, skin Lower injection and intramuscular injection) administration and topical application.In some embodiments, the composition is administered locally to.

The content of bakuchiol composition can be 0.001% to 99.9% weight in the final skin-protection product for PIH Than.In some embodiments, the composition includes 0.1% to 2% Bakuchiol.In other embodiments, described group Close the Bakuchiol that object includes 0.5% or 1.0%.In some embodiments, bakuchiol composition in PIH face cream Amount is 0.5%-1%.Disclosed method includes Bakuchiol including it is a effective amount of orally or topically to give mammalian therapeutic Composition, for complete synthesis or the (or combinations thereof) that is detached from natural source and there is no impurity, especially Furanocoumarin impurities (for example, being less than 500ppm).

The therapeutic agent that the disclosure can be administered locally to by any suitable method well known by persons skilled in the art is used for office Give therapeutic combination in portion.This medication includes but not limited to ointment, gelling agent, lotion or cream base shape Formula or in the form of emulsion, with patches, dressing or facial mask, nonsticking gauze, bandage, cotton swab or cleaning wiping cloth.It can use known This topical application is administered locally to any infected zone by any standard method for local administration.It can be according to medication Therapeutic combination is given with a variety of unit dosage forms.For special delivering mode, treatment can be prepared in the form of above-mentioned excipient Composition.The therapeutic combination of the disclosure can be given to any receptor, preferably give mammal, and more preferably give people.It is special Different administering mode depends on treated morbid state.

No matter which kind of administering mode, according to the specific dosage of approximation re-computation of receptor.Determination has with above-mentioned each preparation Advanced optimizing of being calculated necessary to the suitable dose of the treatment of pass routinely carried out by persons skilled in the art and Without excessively experiment in the working range that they routinely carry out, in particular according to dosage information disclosed herein and inspection.It can Determine that these dosage are used to that suitable dose response data to be combined to determine the dosage used by using the inspection of foundation.One In a little embodiments, including the dosage of the composition of Bakuchiol is the every kg body weights of 0.001mg to 200mg.

In order to illustrate and unrestricted purpose provides the following example.

Embodiment

Embodiment 1

Bakuchiol, psoralen and isopsorapen are quantified by HPLC

By using high pressure liquid chromatography (HPLC) quantified extract of photodiode array detector (HPLC/PDA) Object, fraction, raw materials technology, ingredient and the final amount for preparing Bakuchiol, psoralen and isopsorapen in product.It uses It is 12 minutes that acetonitrile (ACN) or first alcohol and water gradient, which are 36% to 100%CAN times, and the subsequent 100%CAN times are 3 minutes From elution target compound in Luna Phenyl-hexyl columns (250mm × 4.6mm).The detailed HPLC conditions used are explained in table 1 It states.The chromatogram of HPLC separation is shown in FIG. 1.Using commercially available pure Bakuchiol, psoralen and isopsorapen as Plasmid standards for quantitation, based on retention time and the identification of UV peak areas and quantitative objective compound.Bakuchiol, psoralen and different The retention time of psoralen is respectively 18.19 minutes, 7.33 minutes and 7.95 minutes.

Table 1. is used for the HPLC conditions of quantitative Bakuchiol, psoralen and isopsorapen

Embodiment 2

Conventional method for extracting Bakuchiol from Psoralea (PSORALEA) plant

Method ASolvent (100mL) and psoralea corylifolia seed powder (10g) is added to flask, and is shaking manually at room temperature Mixture is shaken 1 hour on bed.Then, mixture by filter and is collected into filtrate.It will be extracted using fresh solvent Journey is repeated once, merging filtrate, removes solvent and under a high vacuum dry residue on a rotary evaporator.

Method BIt is added solvent (50mL) and psoralea corylifolia seed powder (10g) to flask, and mixture is flowed back 40min. Then, solution is filtered and is repeated twice extraction process using fresh solvent.Merging filtrate is simultaneously dry to obtain by evaporation of the solvent Dry extract.

According to above-mentioned extracting process, following solvent extraction samples Plant substance is used：Dichloromethane (DCM), ethyl acetate (EtOAc), acetone, methanol (MeOH), petroleum ether (BP 35-60 DEG C) and petroleum ether (BP 60-90 DEG C).Then, such as embodiment Described in 1, by HPLC analyses come analytical extraction object and plant material.As a result it is illustrated in table 2.

2. various psoralea corylifolia extracts of table quantify

Embodiment 3

Chromatography method for purifying Bakuchiol extract

A variety of chromatography methods are to use for purifying Bakuchiol, the thick solvent extract from thick solvent extract What the method described in embodiment 2 was detached from the seed of psoralea corylifolia.The efficiency of displaying particular column enrichment method, which is used as, not to be had There is the method for the high-purity Bakuchiol of the pollutant of furocoumarin, especially psoralen/Isopsoralen pollutant. Briefly, each empty column shell (1.3cm internal diameters (ID) and 20mL volumes derive from Bio-Rad) is filled up into different media simultaneously Attempt to detach furanocoumarin impurities from Bakuchiol using different solvents elution.Collecting fraction in test tube, (10mL is each Fraction) and the silica gel tlc plate analysis that is unfolded using 20%EtOAc/ petroleum ethers.It is determined based on the solution of n-compound is used Their retention time identification target compound, Bakuchiol, psoralen and isopsorapen.As a result it is explained in table 3 It states.Method described in many tables 3 is used for from synthesis and separation furocoumarin and Bakuchiol in natural source, however for The cost for mass producing this method may not be economically viable.

Table 3. is in the thick extract of psoralea corylifolia from the general introduction of furocoumarin column chromatography for separation Bakuchiol

Embodiment 4

The hydrolysis of the extract detached from the seed of psoralea corylifolia

It will contain from about the hexane extract or CO of the psoralea corylifolia seed of 25% Bakuchiol₂Supercritical fluid extract It is mixed with 1M NaOH solutions.It is at least 1 small that solution, which is heated to 80 DEG C or temperature-time higher than 80 DEG C, in the reaction vessel When.Sub-fraction solution is taken out periodically from flask and is analyzed as described in Example 1 by HPLC.Stop reaction, Show that the peak of psoralen and isopsorapen completely disappears after HPLC analyses.Then, reaction mixture is cooled to room temperature simultaneously Remove water phase.After being washed solution repeatedly using saturation NaCl solution, had using ethyl acetate or the extraction of other organic solvents Machine layer.Organic solution is filtered, washed, is dried and is evaporated has Bakuchiol content not less than 50% and in total to generate There is the brownish red syrup of the psoralen and isopsorapen (Isopsoralen) no more than 100ppm.

Embodiment 5

There is no the antioxidant properties of the bakuchiol composition of furocoumarin

In the laboratories Brunswick, Norton, MA USA evaluations are comprising 57.35% Bakuchiol and are less than in total Natural bakuchiol composition (the batch number UP256- of the psoralen and isopsorapen (Isopsoralen) of 100ppm 0906MP) to the anti-oxidant energy of peroxy radical, hydroxyl free radical, Peroxynitrite, superoxide anion and singlet oxygen Power.According to disclosed technology (Ou, B. et al., J Agric and Food Chem (agricultural with Food Chemistry periodical), 2001, 49(10):4619-4626；Prior, RL. et al., J Agric and Food Chem (agricultural and Food Chemistry periodical), 2005,53:4290-4302) detect the total oxygen radical absorbability of bakuchiol composition.As a result list is aobvious in table 4 Show.

Table 4：Bakutrol^TM(UP256) oxidation resistance properties

Embodiment 6

Evaluation of the bakuchiol composition to the anti-oxidation protection effect of 4-TBP cytotoxicities

Prevent uncle 4- by evaluating it during treatments in 5- days of the compound using concentration under 95% feasibility dosage The aptitude tests of the oxidative stress of butylphenol (4-TBP) induction are less than 100ppm comprising 77.02% Bakuchiol and in total Psoralen and isopsorapen (Isopsoralen) natural bakuchiol composition antioxidant properties.By using The green reactive oxygen species detection kit (InVitrogen) of Image-iT work examines the generation of reactive oxygen species (ROS) to measure Oxidative stress.In the inspection, by carboxyl -2 ', 7 '-dichlorofluorescin diacetate esters are added to the cell stage of culture It is 30 minutes, it diffuses to melanocyte and is 2 ', 7 '-dichlorofluoresceins (DCF) by intracellular ester hydrolysis there, It is reacted with ROS to generate fluorescence DCF.After being treated 5 days using 200 μM or 400 μM of 4-TBP, ROS in melanocyte Generating confirms dose response (that is, being respectively mildly to strong), however the melanocyte of untreated and DMSO treatments does not show ROS is generated.When therapeutic scheme includes test compound, UP256 (Bakuchiol) as shown in Figure 3 shows strong inoxidizability Matter.

Embodiment 7

The tyrosinase inhibitory activity of bakuchiol composition

The tyrosine of two kinds of natural bakuchiol compositions comprising 77.02% Bakuchiol (100% purity) of test Enzyme inhibition activity.Two kinds of substances include the total psoralen and isopsorapen (Isopsoralen) for being less than 100ppm in total.

Use Jones et al., (2002) Pigment.Cell Res.15:The method of 335 reports carries out tyrosinase suppression System is examined.Using this method, by monitoring absorption tracking L-3,4 dihydroxyphenylalanine at 450 nm, the substrate of tyrosinase is converted into DOPA Pigment.Tyrosinase is prepared in 50mM kaliumphosphate buffers under pH 6.8 (examining buffer solution), 2000U/ml and is made With being stored at -20 DEG C with 1ml deciles before.In order to be used for examining, the enzyme solutions of storage are thawed and inspection is used to buffer Liquid is diluted to 200U/ml.The processing solution of 2mM substrates is prepared in examining buffer solution, L-DOPA is used for each inspection.By sample Product are dissolved in 10%DMSO (0.5ml) and are diluted to 5ml using inspection buffer solution.Reaction mixture includes 0.050ml 2mM L- DOPA, 0.050ml 200U/ml Mushroom Tyrosinases and 0.050ml inhibitor.Reaction volume is adjusted using buffer solution is examined To 200 μ l.It tests in 96 hole Falcon, 3097 flat-bottom microtiter plates (Beckton Dickinson, NJ).It uses 1420 multiple labeling counters (Turku, Finland) of WALLAC detect the appearance of dopachrome.It is such as per minute by 450nm Lower absorbance (Δ A₄₅₀) variation detection, from linear enzyme rate determination Mean Speed.Using formula (1) by comparing sample Inhibited by the percentage of the tyrosinase of test sample with the absorbance measurement compareed：

(negative control absorption-sample absorbs)/negative control absorbs × 100 (1)

As shown in figure 4, two kinds of bakuchiol compositions do not show tyrosinase inhibitory activity, and positive control (kojic acid) Show dose reacts tyrosinase and inhibits with 63.9 μM of IC₅₀Value.

Embodiment 8

The preparation of the bakuchiol composition of cosmetic cream, gel and lotion form

As shown below, preparing two kinds with beauty medium or complicated face cream, gel or lotion form includes The natural bakuchiol composition of 86.54% Bakuchiol and 77.02% Bakuchiol.

Preparation A

Preparation B

Formulation C

5. preparation D of table

Embodiment 9

The evaluation of bakuchiol composition security attribute

The Bakuchiol and 77.02% that two kinds include 86.54% is prepared in the form of beauty medium or complicated face cream The natural Bakuchiol combination of Bakuchiol and the in total psoralen and isopsorapen (Isopsoralen) less than 100ppm Their security attribute is tested in object and in vitro model or in people's clinical test.As table 6 is shown, Bakuchiol combination Object does not show eye irritation, contacts sensitization and no phototoxicity without skin irritation, without skin allergy.It is described Composition under the concentration level of wide scope have solid security attribute (Bakuchiol of 20% to 100% weight ratio) and Good Cutaneous permeation property.

Table 6：Bakutrol^TMThe result of safety test

Embodiment 10

There is no the clinical evaluation of the bakuchiol composition of furocoumarin

In the form of beautifying and skin-protecting cream (preparation D, embodiment 8) prepare from the seed of psoralea corylifolia extract and be enriched with and Including 77.02% Bakuchiol and the natural bakuchiol composition (Bakutrol less than the furocoumarin of 100ppm^TM) And it is tested in people's clinical test.Research is tentative, open people's research to evaluate Bakutrol^TMAfter topical application Clinical benefit under 0.5% concentration.Research includes 5 individuals for meeting exclusion/inclusion criteria for evaluating natural Psoralen Benefit of the fat phenol composition for improvement postinflammatory hyperpigmentation (PIH).It is 12 weeks to study the duration.Individual is referred to Show early daily and evening application Bakutrol twice^TM0.5% emulsifiable paste, and be back to position and amount to 9 interviews, including screening is visited Depending on.Evaluation include skin condition (IGA) researcher's net assessment and scytitis after hyperpigmentation (PIH) whole Grade and severity and other relevant skin disease states, including erythema, drying, decortication, oiliness, safety and resistance to It is evaluated by property.Individual questionnaire survey includes related with the use of irritation, skin-friendliness, other products and suncream Safety and compliance sex chromosome mosaicism.In benchmark, the 4th week, the 8th week and the 12nd week shooting photo.It records and analyzes PIH severity From the variation, variation of the PIH grades from benchmark and the successful ratio according to IGA scales of benchmark.Following 6 horizontal serious The PIH grades (0=does not have, and 1=is slight, and 2=is slight, 3=moderates, and 4=is medium serious, and 5=is serious) and PIH of degree infect The gross area of buccal surface analyzed for clinical output.

As shown in figure 5, including Bakutrol in 0.5% time use^TMLocal creams treatment all five individuals have The reduction of at least one grade of PIH severity.Improve in the percentage of 8 weeks lasting facial areas using rear PIH infection More than 50% (referring to Fig. 6).The average percent and absolute scale level of both PIH and its severity improve in Fig. 7 and 8 Middle general introduction.Early in using extract and be enriched with from the seed of psoralea corylifolia and Bakuchiol that include 77.02% and being less than The natural bakuchiol composition of the furocoumarin of 100ppm just realizes being more than for both PIH and severity after 4 weeks 40% or more than a hierarchy level improvement.On the skin of face position of infection PIH substantially reduce it is shown in Fig. 9 It is clearly illustrated in two individual photos.In topical application Bakutrol^TMAfter emulsifiable paste, two individual displays with it is slight and in The gradually improvement of hyperpigmentation (PIH) after the related scytitis of degree acne.

Table 7 outlines compared with the epidemic acne treatment product comprising antiseptic or anti-inflammatory agent or combinations thereof, using not having There is the clinical output of the bakuchiol composition of furocoumarin.Data clearly show the Bakuchiol of not furocoumarin Composition not only improves inflammatory and non-inflammatory lesions count but also significantly improve hyperpigmentation after scytitis.As implemented What example 7 confirmed, tyrosinase is lacked based on it and is inhibited, the PIH benefits derived from bakuchiol composition are unexpected.

The clinical report of 7. Bakutrol and OTC drugs of table is summarized

Embodiment 11

There is no the evaluation of influence of the bakuchiol composition of furocoumarin to reduction postinflammatory hyperpigmentation

Bakuchiol (UP256) emulsifiable paste of 0.5% (wt/wt) of evaluation in double-blind placebo-controlled and positive control research Safety and effect are for treating PIH related with acne.The Psoralen under 0.5% concentration in research evaluation Asia population Fat phenol emulsifiable paste, 2% salicylic acid emulsifiable paste and placebo emulsifiable paste (medium).Participant is instructed to facial application daily twice (AM/PM) emulsifiable paste is studied.Research participant is given application note and provides suncream.

Research object is by more than 18 years old and less than 40 years old and the man in overall physical health that is such as determined by case history Property and female individual composition.18 individuals are collected by Bakuchiol seminar, and 20 individuals are collected by salicylic acid (SAL) seminar, and 19 individuals are collected by placebo seminar.

Investigator and researcher discuss and decide through consultation and such as explicitly defined with acne or the related PIH of other tissue damages (it does not include freckle (Ephilides), solar lentigines freckle (freckle (lentigines)) or chloasma (melisma)).PIH Grade is the measurement (higher number=more serious pigmentation) of hyperpigmented severity.PATIENT POPULATION has PIH grades>3 and acne be slightly to moderate grade 2-3, principal element is related with acne after inflammation is current or inflammation PIH。

Main goal in research：

1. PIH IGA time frames：Benchmark and the 2nd week, the 4th week and the 8th week

2. PIH% distribution time frames：Benchmark and the 2nd week, the 4th week and the 8th week

Using t- test method(s)s or/and variance analysis based on the variation (p from benchmark<0.05) (the p and in other treatment groups< 0.05) PIH effects are evaluated.

By-end：Security evaluation

In benchmark and the evaluation questionnaire for collecting individual in the 2nd week, the 4th week and the 8th week investigation and tolerability evaluations.In benchmark The urine pregnancy test for collecting the women for having reproductive potential with the 8th week.

Data analysis

Collect following data：

Variation of the 1.PIH severity from benchmark；

Variation of the 2.PIH grades from benchmark；With

3. lesion counts the variation from benchmark

As a result:

Data are analyzed in being assessed from investigator in each interview and from the photo that stipulated time point is shot.It is grinding Study carefully before not being mixed by the second investigator group and two individual dermatologists evaluations come from benchmark, the 4th week and the 8th The photo in week is further to confirm that crowd meets standard.Assessment of the data of analysis based on confirmation, the not no photo of second week, Therefore do not include the data of second week in analysis.Table 8 outlines data.

The PIH change of rank of 8. seminar of table

Bakutrol (UP256) group be shown in the 8th week PIH grades (that is, lower PIH grades) from the notable change of benchmark Change (p<0.05).In addition, Bakutrol groups are shown in the significant changes (p on the 8th week placebo<0.05).Data are also shown Bakutrol treatment groups are unique groups (p=0.0083) with time and the notable p value for the treatment of.

The inflammatory acne lesions that 9. all groups of table

Bakutrol (UP256) group is shown in the 4th week and the 8th week significant changes (p from benchmark<0.001). Bakutrol groups are additionally shown in the significant changes (p on the 8th week placebo<0.05).In addition, UP256 is uniquely to be reached at the 8th week The group (57%) more than 50% is reduced to inflammatory lesion.

Bakutrol (UP256) organizes no significant changes in non-inflammatory lesions type (data are not shown)；However, In treatment in the 4th week, salicylate treatment group reached p value in non-inflammatory lesions type<0.05.

Table 10：The general introduction of questionnaire survey and investigator's security evaluation

Conclusion:

As a result be shown in Bakutrol (UP256) emulsifiable paste after only 4 weeks topical applications substantially reduce it is related with acne Postinflammatory hyperpigmentation (PIH).After the 8th week UP256 also substantially reduce inflammatory acne lesions (<0.05) 57%.

Bakutrol (UP256) emulsifiable paste have good security attribute and study participant tolerance it is good.Emulsifiable paste Beauty acceptability be considered acceptable superior or equal to the beauty of their pervious local over the counter (OTC) acne treatments Property.

Embodiment 12

The treatment of inflammatory and non-inflammatory acne lesions

Prepare includes Bakuchiol and salicylic acid combination object.Using composition treatment with inflammatory, non-inflammation The patients with acne of both property or two kinds of lesion.The composition is effective to treatment inflammatory and non-inflammatory lesions, the There is p value when treatment in 4 weeks<0.05.Lesion is reduced to about 10% to about 90%.

It can be by above-mentioned each combination of embodiment to provide other embodiments.Number is referred to and/or applied in specification According to enumerated in table all United States Patent (USP)s, U.S. Patent Application Publication, U.S. Patent application, foreign patent, foreign patent application It is incorporated herein by reference with their entire content with non-patent application.If desired, multiple sides of embodiment can be changed Face is to use each patent, application and disclosed concept to provide other embodiments.It can be according to foregoing detailed description These and other change is carried out to embodiment.In general, in following claims, the term used should not be construed as limiting The right of specific embodiment disclosed in description and claims, and all possible embodiment party should be interpreted as including The full scope for the equivalent that case is given together with such claim.Therefore, claim is not limited by the disclosure.

Claims

1. the composition comprising Bakuchiol or its pharmaceutically acceptable salt prepare for prevent, alleviate, reduce or treat by Purposes in the topical formulations of hyperpigmentation caused by morbid state from inflammatory skin disorders.

2. purposes as described in claim 1, wherein the Bakuchiol is the Bakuchiol of synthesis.

3. purposes as claimed in claim 2, wherein the Bakuchiol of the synthesis is the mixture of stereoisomer.

4. purposes as described in claim 1, wherein the Bakuchiol is from Psoralea, Sassafras, Magnolia or rhizoma atractylodis It is detached in the plant species of category.

5. purposes as described in claim 1 or 4, wherein the Bakuchiol is from entire plant or from seed, stem, tree It is detached in skin, branch, root, root skin, young shoot, flower or other reproductive organs, leaf or other aerial parts or combinations thereof.

6. purposes as claimed in claim 5, wherein the stem includes stem tuber and rhizome.

7. purposes as described in claim 1, wherein the hyperpigmentation from inflammatory skin disorders result from acne, Atopic dermatitis, allergic contact dermatitis, bloch-Siemens syndrome, lichen planus, lupus erythematosus, morphoea, mechanical trauma, electricity From or Non-ionizing radiation, burn, laser or drug therapy, skin infection or combinations thereof.

8. purposes as described in claim 1, wherein the color excessive caused by the morbid state from inflammatory skin disorders It is plain calm for acne spot.

9. purposes as described in claim 1, wherein the composition includes the psoralea corylifolia of 0.001% to 99.9% total weight Phenol and pharmaceutically acceptable, dermatology is acceptable or the acceptable carrier of beauty.

10. purposes as described in claim 1, wherein the composition also includes one kind or mixture of skin whitener, institute It states skin whitener and is selected from quinhydrones, single-benzyl ether, ursin, deoxyarbutin, p methoxy phenol, N- acetyl group -4-S- half Cystamine base phenol, kojic acid, azelaic acid, glycolic, gentianic acid, flavonoids, Aloesin, talan and diphenyl ethylene derivatives, Radix Glycyrrhizae extract, black bearberry extract, mulberry extract, Aloe Vera Gel, glabridin, vitamin C derivatives, vitamin e derivative, Tranexamic acid and its derivative, the biosimulation object of TGF-B protein, centaurcidin, niacinamide, PAR-2 inhibitor, agglutination Element, Neoglycoproteins, resorcinol and its derivative and Nivitol.

11. purposes as claimed in claim 10, wherein the vitamin C derivatives include magnesium ascorbyl phosphate and four hexyls Decyl ascorbic acid.

12. purposes as described in claim 1, wherein the composition also includes one kind or mixture of skin conditioner, institute It states skin conditioner and is selected from 'alpha '-hydroxy acids, salicylic acid, linolenic acid, vitamin A acid, benzoyl peroxide, Guttae Sulfacetamidi Natrici, crin Mycin, erythromycin, dapsone, tetracycline, doxycycline, minocycline, zinc, estrogen and its derivative, antiandrogen, Sulphur, steroids, cortisone, tazarotene, curcumin extract, Arabic gum extract, radix scutellariae extract, Green tea extract and Grape Seed Extract.

13. purposes as described in claim 1, wherein the composition is formulated as in ointment, gel, lotion, emulsifiable paste matrix Emulsion, plaster, dressing or facial mask, nonsticking gauze, bandage, cotton swab or cleaning wiping cloth.